Microfabricated Elastomeric Valve and Pump Systems

ABSTRACT

A method of fabricating an elastomeric structure, comprising: forming a first elastomeric layer on top of a first micromachined mold, the first micromachined mold having a first raised protrusion which forms a first recess extending along a bottom surface of the first elastomeric layer; forming a second elastomeric layer on top of a second micromachined mold, the second micromachined mold having a second raised protrusion which forms a second recess extending along a bottom surface of the second elastomeric layer; bonding the bottom surface of the second elastomeric layer onto a top surface of the first elastomeric layer such that a control channel forms in the second recess between the first and second elastomeric layers; and positioning the first elastomeric layer on top of a planar substrate such that a flow channel forms in the first recess between the first elastomeric layer and the planar substrate.

CROSS-REFERENCE TO RELATED APPLICATIONS

This is a continuation of U.S. patent application Ser. No. 09/724,967,filed Nov. 28, 2000, which is a continuation-in-part of U.S. patentapplication Ser. No. 09/605,520, filed Jun. 27, 2000, which claims thebenefit of the following: U.S. Provisional Patent Application No.60/141,503, filed Jun. 28, 1999, U.S. Provisional Patent Application No.60/147,199, filed Aug. 3, 1999, and U.S. Provisional Patent ApplicationNo. 60/186,856, filed Mar. 3, 2000. The text of these prior provisionalpatent applications is hereby incorporated by reference.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

This invention was made with government support under Grant No. HG01642-02 awarded by the National Institutes of Health. The governmenthas certain rights in the invention.

TECHNICAL FIELD

The present invention relates to microfabricated structures and methodsfor producing microfabricated structures, and to microfabricated systemsfor regulating fluid-flow.

BACKGROUND OF THE INVENTION

Various approaches to designing micro-fluidic pumps and valves have beenattempted. Unfortunately, each of these approaches suffers from its ownlimitations.

The two most common methods of producing microelectromechanical (MEMS)structures such as pumps and valves are silicon-based bulkmicro-machining (which is a subtractive fabrication method wherebysingle crystal silicon is lithographically patterned and then etched toform three-dimensional structures), and surface micro-machining (whichis an additive method where layers of semiconductor-type materials suchas polysilicon, silicon nitride, silicon dioxide, and various metals aresequentially added and patterned to make three-dimensional structures).

A limitation of the first approach of silicon-based micro-machining isthat the stiffness of the semiconductor materials used necessitates highactuation forces, which in turn result in large and complex designs. Infact, both bulk and surface micro-machining methods are limited by thestiffness of the materials used. In addition, adhesion between variouslayers of the fabricated device is also a problem. For example, in bulkmicro-machining, wafer bonding techniques must be employed to createmultilayer structures. On the other hand, when surface micro-machining,thermal stresses between the various layers of the device limits thetotal device thickness, often to approximately 20 microns. Using eitherof the above methods, clean room fabrication and careful quality controlare required.

SUMMARY OF THE INVENTION

The present invention sets forth systems for fabricating and operatingmicrofabricated structures such as on/off valves, switching valves, andpumps e.g. made out of various layers of elastomer bonded together. Thepresent structures and methods are ideally suited for controlling andchanneling fluid movement, but are not so limited.

In a preferred aspect, the present invention uses a multilayer softlithography process to build integrated (i.e.: monolithic)microfabricated elastomeric structures.

Advantages of fabricating the present structures by binding togetherlayers of soft elastomeric materials include the fact that the resultingdevices are reduced by more than two orders of magnitude in size ascompared to silicon-based devices. Further advantages of rapidprototyping, ease of fabrication, and biocompatability are alsoachieved.

In preferred aspects of the invention, separate elastomeric layers arefabricated on top of micromachined molds such that recesses are formedin each of the various elastomeric layers. By bonding these variouselastomeric layers together, the recesses extending along the variouselastomeric layers form flow channels and control lines through theresulting monolithic, integral elastomeric structure. In various aspectsof the invention, these flow channels and control lines which are formedin the elastomeric structure can be actuated to function as micro-pumpsand micro-valves, as will be explained.

In further optional aspects of the invention, the monolithic elastomericstructure is sealed onto the top of a planar substrate, with flowchannels being formed between the surface of the planar substrate andthe recesses which extend along the bottom surface of the elastomericstructure.

In one preferred aspect, the present monolithic elastomeric structuresare constructed by bonding together two separate layers of elastomerwith each layer first being separately cast from a micromachined mold.Preferably, the elastomer used is a two-component addition cure materialin which the bottom elastomeric layer has an excess of one component,while the top elastomeric layer has an excess of another component. Inan exemplary embodiment, the elastomer used is silicone rubber. Twolayers of elastomer are cured separately. Each layer is separately curedbefore the top layer is positioned on the bottom layer. The two layersare then bonded together. Each layer preferably has an excess of one ofthe two components, such that reactive molecules remain at the interfacebetween the layers. The top layer is assembled on top of the bottomlayer and heated. The two layers bond irreversibly such that thestrength of the interface approaches or equals the strength of the bulkelastomer. This creates a monolithic three-dimensional patternedstructure composed entirely of two layers of bonded together elastomer.Additional layers may be added by simply repeating the process, whereinnew layers, each having a layer of opposite “polarity” are cured, andthereby bonded together.

In a second preferred aspect, a first photoresist layer is deposited ontop of a first elastomeric layer. The first photoresist layer is thenpatterned to leave a line or pattern of lines of photoresist on the topsurface of the first elastomeric layer. Another layer of elastomer isthen added and cured, encapsulating the line or pattern of lines ofphotoresist. A second photoresist layer is added and patterned, andanother layer of elastomer added and cured, leaving line and patterns oflines of photoresist encapsulated in a monolithic elastomer structure.This process may be repeated to add more encapsulated patterns andelastomer layers. Thereafter, the photoresist is removed leaving flowchannel(s) and control line(s) in the spaces which had been occupied bythe photoresist. This process may be repeated to create elastomerstructures having a multitude of layers.

An advantage of patterning moderate sized features (>/=10 microns) usinga photoresist method is that a high resolution transparency film can beused as a contact mask. This allows a single researcher to design,print, pattern the mold, and create a new set of cast elastomer devices,typically all within 24 hours.

A further advantage of either above embodiment of the present inventionis that due to its monolithic or integral nature, (i.e., all the layersare composed of the same material) is that interlayer adhesion failuresand thermal stress problems are completely avoided.

Further advantages of the present invention's preferred use of asilicone rubber or elastomer such as RTV 615 manufactured by GeneralElectric, is that it is transparent to visible light, making amultilayer optical trains possible, thereby allowing opticalinterrogation of various channels or chambers in the microfluidicdevice. As appropriately shaped elastomer layers can serve as lenses andoptical elements, bonding of layers allows the creation of multilayeroptical trains. In addition, GE RTV 615 elastomer is biocompatible.Being soft, closed valves form a good seal even if there are smallparticulates in the flow channel. Silicone rubber is also bio-compatibleand inexpensive, especially when compared with a single crystal silicon.

Monolithic elastomeric valves and pumps also avoid many of the practicalproblems affecting flow systems based on electro-osmotic flow.Typically, electro-osmotic flow systems suffer from bubble formationaround the electrodes and the flow is strongly dependent on thecomposition of the flow medium. Bubble formation seriously restricts theuse of electro-osmotic flow in microfluidic devices, making it difficultto construct functioning integrated devices. The magnitude of flow andeven its direction typically depends in a complex fashion on ionicstrength and type, the presence of surfactants and the charge on thewalls of the flow channel. Moreover, since electrolysis is taking placecontinuously, the eventual capacity of buffer to resist pH changes mayalso be reached. Furthermore, electro-osmotic flow always occurs incompetition with electrophoresis. As different molecules may havedifferent electrophoretic mobilities, unwanted electrophoreticseparation may occur in the electro-osmotic flow. Finally,electro-osmotic flow can-not easily be used to stop flow, haltdiffusion, or to balance pressure differences.

A further advantage of the present monolithic elastomeric valve and pumpstructures are that they can be actuated at very high speeds. Forexample, the present inventors have achieved a response time for a valvewith aqueous solution therein on the order of one millisecond, such thatthe valve opens and closes at speeds approaching or exceeding 100 Hz. Inparticular, a non-exclusive list of ranges of cycling speeds for theopening and closing of the valve structure include between about 0.001and 10000 ms, between about 0.01 and 1000 ms, between about 0.1 and 100ms, and between about 1 and 10 ms. The cycling speeds depend upon thecomposition and structure of a valve used for a particular applicationand the method of actuation, and thus cycling speeds outside of thelisted ranges would fall within the scope of the present invention.

Further advantages of the present pumps and valves are that their smallsize makes them fast and their softness contributes to making themdurable. Moreover, as they close linearly with differential appliedpressure, this linear relationship allows fluid metering and valveclosing in spite of high back pressures.

In various aspects of the invention, a plurality of flow channels passthrough the elastomeric structure with a second flow channel extendingacross and above a first flow channel. In this aspect of the invention,a thin membrane of elastomer separates the first and second flowchannels. As will be explained, downward movement of this membrane (dueto the second flow channel being pressurized or the membrane beingotherwise actuated) will cut off flow passing through the lower flowchannel.

In optional preferred aspects of the present systems, a plurality ofindividually addressable valves are formed connected together in anelastomeric structure and are then activated in sequence such thatperistaltic pumping is achieved. More complex systems includingnetworked or multiplexed control systems, selectably addressable valvesdisposed in a grid of valves, networked or multiplexed reaction chambersystems and biopolymer synthesis systems are also described.

One embodiment of a microfabricated elastomeric structure in accordancewith the present invention comprises an elastomeric block formed withfirst and second microfabricated recesses therein, a portion of theelastomeric block deflectable when the portion is actuated.

One embodiment of a method of microfabricating an elastomeric structurecomprises the steps of microfabricating a first elastomeric layer,microfabricating a second elastomeric layer; positioning the secondelastomeric layer on top of the first elastomeric layer, and bonding abottom surface of the second elastomeric layer onto a top surface of thefirst elastomeric layer.

A first alternative embodiment of a method of microfabricating anelastomeric structure comprises the steps of forming a first elastomericlayer on top of a first micromachined mold, the first micromachined moldhaving at least one first raised protrusion which forms at least onefirst channel in the bottom surface of the first elastomeric layer. Asecond elastomeric layer is formed on top of a second micromachinedmold, the second micromachined mold having at least one second raisedprotrusion which forms at least one second channel in the bottom surfaceof the second elastomeric layer. The bottom surface of the secondelastomeric layer is bonded onto a top surface of the first elastomericlayer such that the at least one second channel is enclosed between thefirst and second elastomeric layers.

A second alternative embodiment of a method of microfabricating anelastomeric structure in accordance with the present invention comprisesthe steps of forming a first elastomeric layer on top of a substrate,curing the first elastomeric layer, and depositing a first sacrificiallayer on the top surface of the first elastomeric layer. A portion ofthe first sacrificial layer is removed such that a first pattern ofsacrificial material remains on the top surface of the first elastomericlayer. A second elastomeric layer is formed over the first elastomericlayer thereby encapsulating the first pattern of sacrificial materialbetween the first and second elastomeric layers. The second elastomericlayer is cured and then sacrificial material is removed thereby formingat least one first recess between the first and second layers ofelastomer.

An embodiment of a method of actuating an elastomeric structure inaccordance with the present invention comprises providing an elastomericblock formed with first and second microfabricated recesses therein, thefirst and second microfabricated recesses being separated by a portionof the structure which is deflectable into either of the first or secondrecesses when the other of the first and second recesses. One of therecesses is pressurized such that the portion of the elastomericstructure separating the second recess from the first recess isdeflected into the other of the two recesses.

In other optional preferred aspects, magnetic or conductive materialscan be added to make layers of the elastomer magnetic or electricallyconducting, thus enabling the creation of all elastomer electromagneticdevices.

BRIEF DESCRIPTION OF THE DRAWINGS

Part I—FIGS. 1 to 7A illustrate successive steps of a first method offabricating the present invention, as follows:

FIG. 1 is an illustration of a first elastomeric layer formed on top ofa micromachined mold.

FIG. 2 is an illustration of a second elastomeric layer formed on top ofa micromachined mold.

FIG. 3 is an illustration of the elastomeric layer of FIG. 2 removedfrom the micromachined mold and positioned over the top of theelastomeric layer of FIG. 1.

FIG. 4 is an illustration corresponding to FIG. 3, but showing thesecond elastomeric layer positioned on top of the first elastomericlayer.

FIG. 5 is an illustration corresponding to FIG. 4, but showing the firstand second elastomeric layers bonded together.

FIG. 6 is an illustration corresponding to FIG. 5, but showing the firstmicromachine mold removed and a planar substrate positioned in itsplace.

FIG. 7A is an illustration corresponding to FIG. 6, but showing theelastomeric structure sealed onto the planar substrate.

FIG. 7B is a front sectional view corresponding to FIG. 7A, showing anopen flow channel.

FIGS. 7C-7G are illustrations showing steps of a method for forming anelastomeric structure having a membrane formed from a separateelastomeric layer.

Part II—FIG. 7H show the closing of a first flow channel by pressurizinga second flow channel, as follows:

FIG. 7H corresponds to FIG. 7A, but shows a first flow channel closed bypressurization in second flow channel.

Part III—FIGS. 8 to 18 illustrate successive steps of a second method offabricating the present invention, as follows:

FIG. 8 is an illustration of a first elastomeric layer deposited on aplanar substrate.

FIG. 9 is an illustration showing a first photoresist layer deposited ontop of the first elastomeric layer of FIG. 8.

FIG. 10 is an illustration showing the system of FIG. 9, but with aportion of the first photoresist layer removed, leaving only a firstline of photoresist.

FIG. 11 is an illustration showing a second elastomeric layer applied ontop of the first elastomeric layer over the first line of photoresist ofFIG. 10, thereby encasing the photoresist between the first and secondelastomeric layers.

FIG. 12 corresponds to FIG. 11, but shows the integrated monolithicstructure produced after the first and second elastomer layers have beenbonded together.

FIG. 13 is an illustration showing a second photoresist layer depositedon top of the integral elastomeric structure of FIG. 12.

FIG. 14 is an illustration showing the system of FIG. 13, but with aportion of the second photoresist layer removed, leaving only a secondline of photoresist.

FIG. 15 is an illustration showing a third elastomer layer applied ontop of the second elastomeric layer and over the second line ofphotoresist of FIG. 14, thereby encapsulating the second line ofphotoresist between the elastomeric structure of FIG. 12 and the thirdelastomeric layer.

FIG. 16 corresponds to FIG. 15, but shows the third elastomeric layercured so as to be bonded to the monolithic structure composed of thepreviously bonded first and second elastomer layers.

FIG. 17 corresponds to FIG. 16, but shows the first and second lines ofphotoresist removed so as to provide two perpendicular overlapping, butnot intersecting, flow channels passing through the integratedelastomeric structure.

FIG. 18 is an illustration showing the system of FIG. 17, but with theplanar substrate thereunder removed.

Part IV—FIGS. 19 and 20 show further details of different flow channelcross-sections, as follows:

FIG. 19 shows a rectangular cross-section of a first flow channel.

FIG. 20 shows the flow channel cross section having a curved uppersurface.

Part V—FIGS. 21 to 24 show experimental results achieved by preferredembodiments of the present microfabricated valve:

FIG. 21 illustrates valve opening vs. applied pressure for various flowchannels.

FIG. 22 illustrates time response of a 100 μm×100 μm×10 μm RTVmicrovalve.

Part VI—FIGS. 23A to 33 show various microfabricated structures,networked together according to aspects of the present invention:

FIG. 23A is a top schematic view of an on/off valve.

FIG. 23B is a sectional elevation view along line 23B-23B in FIG. 23A.

FIG. 24 is a top schematic view of a peristaltic pumping system.

FIG. 24B is a sectional elevation view along line 24B-24B in FIG. 24A.

FIG. 25 is a graph showing experimentally achieved pumping rates vs.frequency for an embodiment of the peristaltic pumping system of FIG.24.

FIG. 26A is a top schematic view of one control line actuating multipleflow lines simultaneously.

FIG. 26B is a sectional elevation view along line 26B-26B in FIG. 26A.

FIG. 27 is a schematic illustration of a multiplexed system adapted topermit flow through various channels.

FIG. 28A is a plan view of a flow layer of an addressable reactionchamber structure.

FIG. 28B is a bottom plan view of a control channel layer of anaddressable reaction chamber structure.

FIG. 28C is an exploded perspective view of the addressable reactionchamber structure formed by bonding the control channel layer of FIG.28B to the top of the flow layer of FIG. 28A.

FIG. 28D is a sectional elevation view corresponding to FIG. 28C, takenalong line 28D-28D in FIG. 28C.

FIG. 29 is a schematic of a system adapted to selectively direct fluidflow into any of an array of reaction wells.

FIG. 30 is a schematic of a system adapted for selectable lateral flowbetween parallel flow channels.

FIG. 31A is a bottom plan view of first layer (i.e.: the flow channellayer) of elastomer of a switchable flow array.

FIG. 31B is a bottom plan view of a control channel layer of aswitchable flow array.

FIG. 31C shows the alignment of the first layer of elastomer of FIG. 31Awith one set of control channels in the second layer of elastomer ofFIG. 31B.

FIG. 31D also shows the alignment of the first layer of elastomer ofFIG. 31A with the other set of control channels in the second layer ofelastomer of FIG. 31B.

FIG. 32 is a schematic of an integrated system for biopolymer synthesis.

FIG. 33 is a schematic of a further integrated system for biopolymersynthesis.

FIG. 34 is an optical micrograph of a section of a test structure havingseven layers of elastomer bonded together.

FIGS. 35A-35D show the steps of one embodiment of a method forfabricating an elastomer layer having a vertical via formed therein.

FIG. 36 shows one embodiment of a sorting apparatus in accordance withthe present invention.

FIG. 37 shows an embodiment of an apparatus for flowing process gasesover a semiconductor wafer in accordance with the present invention.

FIG. 38 shows an exploded view of one embodiment of a micro-mirror arraystructure in accordance with the present invention.

FIG. 39 shows a perspective view of a first embodiment of a refractivedevice in accordance with the present invention.

FIG. 40 shows a perspective view of a second embodiment of a refractivedevice in accordance with the present invention.

FIG. 41 shows a perspective view of a third embodiment of a refractivedevice in accordance with the present invention.

FIGS. 42A-42J show views of one embodiment of a normally-closed valvestructure in accordance with the present invention.

FIG. 43 shows a plan view of one embodiment of a device for performingseparations in accordance with the present invention.

FIGS. 44A-44D show plan views illustrating operation of one embodimentof a cell pen structure in accordance with the present invention.

FIGS. 45A-45B show plan and cross-sectional views illustrating operationof one embodiment of a cell cage structure in accordance with thepresent invention.

FIGS. 46A-46B show cross-sectional views illustrating operation of oneembodiment of a cell grinder structure in accordance with the presentinvention.

FIG. 47 shows a plan view of one embodiment of a pressure oscillatorstructure in accordance with the present invention.

FIGS. 48A and 48B show plan views illustrating operation of oneembodiment of a side-actuated valve structure in accordance with thepresent invention.

FIG. 49 plots Young's modulus versus percentage dilution of GE RTV 615elastomer with GE SF96-50 silicone fluid.

FIG. 50 shows a cross-sectional view of a structure in whichchannel-bearing faces are placed into contact to form a larger-sizedchannel.

FIG. 51 shows a cross-sectional view of a structure in which non-channelbearing faces are placed into contact and then sandwiched between twosubstrates.

FIGS. 52A-52C show cross-sectional views of the steps for constructing abridging structure. FIG. 52D shows a plan view of the bridgingstructure.

FIG. 53 shows a cross-sectional view of one embodiment of a compositestructure in accordance with the present invention.

FIG. 54 shows a cross-sectional view of one embodiment of a compositestructure in accordance with the present invention.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

The present invention comprises a variety of microfabricated elastomericstructures which may be used as pumps or valves. Methods of fabricatingthe preferred elastomeric structures are also set forth.

Methods of Fabricating the Present Invention:

Two exemplary methods of fabricating the present invention are providedherein. It is to be understood that the present invention is not limitedto fabrication by one or the other of these methods. Rather, othersuitable methods of fabricating the present microstructures, includingmodifying the present methods, are also contemplated.

FIGS. 1 to 7B illustrate sequential steps of a first preferred method offabricating the present microstructure, (which may be used as a pump orvalve). FIGS. 8 to 18 illustrate sequential steps of a second preferredmethod of fabricating the present microstructure, (which also may beused as a pump or valve).

As will be explained, the preferred method of FIGS. 1 to 7B involvesusing pre-cured elastomer layers which are assembled and bonded.Conversely, the preferred method of FIGS. 8 to 18 involves curing eachlayer of elastomer “in place.” In the following description “channel”refers to a recess in the elastomeric structure which can contain a flowof fluid or gas.

The First Exemplary Method:

Referring to FIG. 1, a first micro-machined mold 10 is provided.Micro-machined mold 10 may be fabricated by a number of conventionalsilicon processing methods, including but not limited tophotolithography, ion-milling, and electron beam lithography.

As can be seen, micro-machined mold 10 has a raised line or protrusion11 extending therealong. A first elastomeric layer 20 is cast on top ofmold 10 such that a first recess 21 will be formed in the bottom surfaceof elastomeric layer 20, (recess 21 corresponding in dimension toprotrusion 11), as shown.

As can be seen in FIG. 2, a second micro-machined mold 12 having araised protrusion 13 extending therealong is also provided. A secondelastomeric layer 22 is cast on top of mold 12, as shown, such that arecess 23 will be formed in its bottom surface corresponding to thedimensions of protrusion 13.

As can be seen in the sequential steps illustrated in FIGS. 3 and 4,second elastomeric layer 22 is then removed from mold 12 and placed ontop of first elastomeric layer 20. As can be seen, recess 23 extendingalong the bottom surface of second elastomeric layer 22 will form a flowchannel 32.

Referring to FIG. 5, the separate first and second elastomeric layers 20and 22 FIG. 4 are then bonded together to form an integrated (i.e.:monolithic) elastomeric structure 24.

As can been seen in the sequential step of FIGS. 6 and 7A, elastomericstructure 24 is then removed from mold 10 and positioned on top of aplanar substrate 14. As can be seen in FIGS. 7A and 7B, when elastomericstructure 24 has been sealed at its bottom surface to planar substrate14, recess 21 will form a flow channel 30.

The present elastomeric structures form a reversible hermetic seal withnearly any smooth planar substrate. An advantage to forming a seal thisway is that the elastomeric structures may be peeled up, washed, andre-used. In preferred aspects, planar substrate 14 is glass. A furtheradvantage of using glass is that glass is transparent, allowing opticalinterrogation of elastomer channels and reservoirs. Alternatively, theelastomeric structure may be bonded onto a flat elastomer layer by thesame method as described above, forming a permanent and high-strengthbond. This may prove advantageous when higher back pressures are used.

As can be seen in FIGS. 7A and 7B, flow channels 30 and 32 arepreferably disposed at an angle to one another with a small membrane 25of substrate 24 separating the top of flow channel 30 from the bottom offlow channel 32.

In preferred aspects, planar substrate 14 is glass. An advantage ofusing glass is that the present elastomeric structures may be peeled up,washed and reused. A further advantage of using glass is that opticalsensing may be employed. Alternatively, planar substrate 14 may be anelastomer itself, which may prove advantageous when higher backpressures are used.

The method of fabrication just described may be varied to form astructure having a membrane composed of an elastomeric materialdifferent than that forming the walls of the channels of the device.This variant fabrication method is illustrated in FIGS. 7C-7G.

Referring to FIG. 7C, a first micro-machined mold 10 is provided.Micro-machined mold 10 has a raised line or protrusion 11 extendingtherealong. In FIG. 7D, first elastomeric layer 20 is cast on top offirst micro-machined mold 10 such that the top of the first elastomericlayer 20 is flush with the top of raised line or protrusion 11. This maybe accomplished by carefully controlling the volume of elastomericmaterial spun onto mold 10 relative to the known height of raised line11. Alternatively, the desired shape could be formed by injectionmolding.

In FIG. 7E, second micro-machined mold 12 having a raised protrusion 13extending therealong is also provided. Second elastomeric layer 22 iscast on top of second mold 12 as shown, such that recess 23 is formed inits bottom surface corresponding to the dimensions of protrusion 13.

In FIG. 7F, second elastomeric layer 22 is removed from mold 12 andplaced on top of third elastomeric layer 222. Second elastomeric layer22 is bonded to third elastomeric layer 20 to form integral elastomericblock 224 using techniques described in detail below. At this point inthe process, recess 23 formerly occupied by raised line 13 will formflow channel 23.

In FIG. 7G, elastomeric block 224 is placed on top of firstmicro-machined mold 10 and first elastomeric layer 20. Elastomeric blockand first elastomeric layer 20 are then bonded together to form anintegrated (i.e.: monolithic) elastomeric structure 24 having a membranecomposed of a separate elastomeric layer 222.

When elastomeric structure 24 has been sealed at its bottom surface to aplanar substrate in the manner described above in connection with FIG.7A, the recess formerly occupied by raised line 11 will form flowchannel 30.

The variant fabrication method illustrated above in conjunction withFIGS. 7C-7G offers the advantage of permitting the membrane portion tobe composed of a separate material than the elastomeric material of theremainder of the structure. This is important because the thickness andelastic properties of the membrane play a key role in operation of thedevice. Moreover, this method allows the separate elastomer layer toreadily be subjected to conditioning prior to incorporation into theelastomer structure. As discussed in detail below, examples ofpotentially desirable condition include the introduction of magnetic orelectrically conducting species to permit actuation of the membrane,and/or the introduction of dopant into the membrane in order to alterits elasticity.

While the above method is illustrated in connection with forming variousshaped elastomeric layers formed by replication molding on top of amicromachined mold, the present invention is not limited to thistechnique. Other techniques could be employed to form the individuallayers of shaped elastomeric material that are to be bonded together.For example, a shaped layer of elastomeric material could be formed bylaser cutting or injection molding, or by methods utilizing chemicaletching and/or sacrificial materials as discussed below in conjunctionwith the second exemplary method.

The Second Exemplary Method:

A second exemplary method of fabricating an elastomeric structure whichmay be used as a pump or valve is set forth in the sequential stepsshown in FIGS. 8-18.

In this aspect of the invention, flow and control channels are definedby first patterning photoresist on the surface of an elastomeric layer(or other substrate, which may include glass) leaving a raised linephotoresist where a channel is desired. Next, a second layer ofelastomer is added thereover and a second photoresist is patterned onthe second layer of elastomer leaving a raised line photoresist where achannel is desired. A third layer of elastomer is deposited thereover.Finally, the photoresist is removed by dissolving it out of theelastomer with an appropriate solvent, with the voids formed by removalof the photoresist becoming the flow channels passing through thesubstrate.

Referring first to FIG. 8, a planar substrate 40 is provided. A firstelastomeric layer 42 is then deposited and cured on top of planarsubstrate 40. Referring to FIG. 9, a first photoresist layer 44A is thendeposited over the top of elastomeric layer 42. Referring to FIG. 10, aportion of photoresist layer 44A is removed such that only a first lineof photoresist 44B remains as shown. Referring to FIG. 11, a secondelastomeric layer 46 is then deposited over the top of first elastomericlayer 42 and over the first line of photoresist 44B as shown, therebyencasing first line of photoresist 44B between first elastomeric layer42 and second elastomeric layer 46. Referring to FIG. 12, elastomericlayers 46 is then cured on layer 42 to bond the layers together to forma monolithic elastomeric substrate 45.

Referring to FIG. 13, a second photoresist layer 48A is then depositedover elastomeric structure 45. Referring to FIG. 14, a portion of secondphotoresist layer 48A is removed, leaving only a second photoresist line48B on top of elastomeric structure 45 as shown. Referring to FIG. 15, athird elastomeric layer 50 is then deposited over the top of elastomericstructure 45 (comprised of second elastomeric layer 42 and first line ofphotoresist 44B) and second photoresist line 48B as shown, therebyencasing the second line of photoresist 48B between elastomericstructure 45 and third elastomeric layer 50.

Referring to FIG. 16, third elastomeric layer 50 and elastomericstructure 45 (comprising first elastomeric layer 42 and secondelastomeric layer 46 bonded together) is then bonded together forming amonolithic elastomeric structure 47 having photoresist lines 44B and 48Bpassing therethrough as shown. Referring to FIG. 17, photoresist lines44B, 48B are then removed (for example, by an solvent) such that a firstflow channel 60 and a second flow channel 62 are provided in theirplace, passing through elastomeric structure 47 as shown. Lastly,referring to FIG. 18, planar substrate 40 can be removed from the bottomof the integrated monolithic structure.

The method described in FIGS. 8-18 fabricates a patterned elastomerstructure utilizing development of photoresist encapsulated withinelastomer material. However, the methods in accordance with the presentinvention are not limited to utilizing photoresist. Other materials suchas metals could also serve as sacrificial materials to be removedselective to the surrounding elastomer material, and the method wouldremain within the scope of the present invention. For example, asdescribed in detail below in connection with FIGS. 35A-35D, gold metalmay be etched selective to RTV 615 [trade] elastomer utilizing theappropriate chemical mixture.

Preferred Layer and Channel Dimensions:

Microfabricated refers to the size of features of an elastomericstructure fabricated in accordance with an embodiment of the presentinvention. In general, variation in at least one dimension ofmicrofabricated structures is controlled to the micron level, with atleast one dimension being microscopic (i.e. below 1000 μm).Microfabrication typically involves semiconductor or MEMS fabricationtechniques such as photolithography and spincoating that are designedfor to produce feature dimensions on the microscopic level, with atleast some of the dimension of the microfabricated structure requiring amicroscope to reasonably resolve/image the structure.

In preferred aspects, flow channels 30, 32, 60 and 62 preferably havewidth-to-depth ratios of about 10:1. A non-exclusive list of otherranges of width-to-depth ratios in accordance with embodiments of thepresent invention is 0.1:1 to 100:1, more preferably 1:1 to 50:1, morepreferably 2:1 to 20:1, and most preferably 3:1 to 15:1. In an exemplaryaspect, flow channels 30, 32, 60 and 62 have widths of about 1 to 1000microns. A non-exclusive list of other ranges of widths of flow channelsin accordance with embodiments of the present invention is 0.01 to 1000microns, more preferably 0.05 to 1000 microns, more preferably 0.2 to500 microns, more preferably 1 to 250 microns, and most preferably 10 to200 microns. Exemplary channel widths include 0.1 μm, 1 μm, 2 μm, 5 μm,10 μm, 20 μm, 30 μm, 40 μm, 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, 100 μm,110 μm, 120 μm, 130 μm, 140 μm, 150 μm, 160 μm, 170 μm, 180 μm, 190 μm,200 μm, 210 μm, 220 μm, 230 μm, 240 μm, and 250 μm.

Flow channels 30, 32, 60, and 62 have depths of about 1 to 100 microns.A non-exclusive list of other ranges of depths of flow channels inaccordance with embodiments of the present invention is 0.01 to 1000microns, more preferably 0.05 to 500 microns, more preferably 0.2 to 250microns, and more preferably 1 to 100 microns, more preferably 2 to 20microns, and most preferably 5 to 10 microns. Exemplary channel depthsinclude including 0.01 μm, 0.02 μm, 0.05 μm, 0.1 μm, 0.2 μm, 0.5 μm, 1μm, 2 μm, 3 μm, 4 μm, 5 μm, 7.5 μm, 10 μm, 12.5 μm, 15 μm, 17.5 μm, 20μm, 22.5 μm, 25 μm, 30 μm, 40 μm, 50 μm, 75 μm, 100 μm, 150 μm, 200 μm,and 250 μm.

The flow channels are not limited to these specific dimension ranges andexamples given above, and may vary in width in order to affect themagnitude of force required to deflect the membrane as discussed atlength below in conjunction with FIG. 27. For example, extremely narrowflow channels having a width on the order of 0.01 μm may be useful inoptical and other applications, as discussed in detail below.Elastomeric structures which include portions having channels of evengreater width than described above are also contemplated by the presentinvention, and examples of applications of utilizing such wider flowchannels include fluid reservoir and mixing channel structures.

Elastomeric layer 22 may be cast thick for mechanical stability. In anexemplary embodiment, layer 22 is 50 microns to several centimetersthick, and more preferably approximately 4 mm thick. A non-exclusivelist of ranges of thickness of the elastomer layer in accordance withother embodiments of the present invention is between about 0.1 micronto 10 cm, 1 micron to 5 cm, 10 microns to 2 cm, 100 microns to 10 mm.

Accordingly, membrane 25 of FIG. 7B separating flow channels 30 and 32has a typical thickness of between about 0.01 and 1000 microns, morepreferably 0.05 to 500 microns, more preferably 0.2 to 250, morepreferably 1 to 100 microns, more preferably 2 to 50 microns, and mostpreferably 5 to 40 microns. As such, the thickness of elastomeric layer22 is about 100 times the thickness of elastomeric layer 20. Exemplarymembrane thicknesses include 0.01 μm, 0.02 μm, 0.03 μm, 0.05 μm, 0.1 μm,0.2 μm, 0.3 μm, 0.5 μm, 1 μm, 2 μm, 3 μm, 5 μm, 7.5 μm, 10 μm, 12.5 μm,15 μm, 17.5 μm, 20 μm, 22.5 μm, 25 μm, 30 μm, 40 μm, 50 μm, 75 μm, 100μm, 150 μm, 200 μm, 250 μm, 300 μm, 400 μm, 500 μm, 750 μm, and 1000 μm

Similarly, first elastomeric layer 42 may have a preferred thicknessabout equal to that of elastomeric layer 20 or 22; second elastomericlayer 46 may have a preferred thickness about equal to that ofelastomeric layer 20; and third elastomeric layer 50 may have apreferred thickness about equal to that of elastomeric layer 22.

Multilayer Soft Lithography Construction Techniques and Materials: SoftLithographic Bonding:

Preferably, elastomeric layers 20 and 22 (or elastomeric layers 42, 46and 50) are bonded together chemically, using chemistry that isintrinsic to the polymers comprising the patterned elastomer layers.Most preferably, the bonding comprises two component “addition cure”bonding.

In a preferred aspect, the various layers of elastomer are boundtogether in a heterogenous bonding in which the layers have a differentchemistry. Alternatively, a homogenous bonding may be used in which alllayers would be of the same chemistry. Thirdly, the respective elastomerlayers may optionally be glued together by an adhesive instead. In afourth aspect, the elastomeric layers may be thermoset elastomers bondedtogether by heating.

In one aspect of homogeneous bonding, the elastomeric layers arecomposed of the same elastomer material, with the same chemical entityin one layer reacting with the same chemical entity in the other layerto bond the layers together. In one embodiment, bonding between polymerchains of like elastomer layers may result from activation of acrosslinking agent due to light, heat, or chemical reaction with aseparate chemical species.

Alternatively in a heterogeneous aspect, the elastomeric layers arecomposed of different elastomeric materials, with a first chemicalentity in one layer reacting with a second chemical entity in anotherlayer. In one exemplary heterogeneous aspect, the bonding process usedto bind respective elastomeric layers together may comprise bondingtogether two layers of RTV 615 [trade] silicone. RTV 615 [trade]silicone is a two-part addition-cure silicone rubber. Part A containsvinyl groups and catalyst; part B contains silicon hydride (Si—H)groups. The conventional ratio for RTV 615 [trade] is 10A:1B. Forbonding, one layer may be made with 30A:1B (i.e. excess vinyl groups)and the other with 3A:1B (i.e. excess Si—H groups). Each layer is curedseparately. When the two layers are brought into contact and heated atelevated temperature, they bond irreversibly forming a monolithicelastomeric substrate.

In an exemplary aspect of the present invention, elastomeric structuresare formed utilizing Sylgard [®] 182, 184 or 186, or aliphatic urethanediacrylates such as (but not limited to) Ebecryl 270 [®] or In 245 [®]from UCB Chemical [®].

In one embodiment in accordance with the present invention, two-layerelastomeric structures were fabricated from pure acrylated Urethane Ebe270 [®]. A thin bottom layer was spin coated at 8000 rpm for 15 secondsat 170° C. The top and bottom layers were initially cured underultraviolet light for 10 minutes under nitrogen utilizing a Model ELC500 [trade] device manufactured by ELECTROLITE [trade] corporation. Theassembled layers were then cured for an additional 30 minutes. Reactionwas catalyzed by a 0.5% vol/vol mixture of Irgacure [®] 500 manufacturedby CIBA-GEIGY [®] Chemicals. The resulting elastomeric materialexhibited moderate elasticity and adhesion to glass.

In another embodiment in accordance with the present invention,two-layer elastomeric structures were fabricated from a combination of25% Ebe 270/50% Irr245/25% isopropyl alcohol for a thin bottom layer,and pure acrylated URETHANE EBE 270 as a top layer. The thin bottomlayer was initially cured for 5 min, and the top layer initially curedfor 10 minutes, under ultraviolet light under nitrogen utilizing a ModelELC 500 device manufactured by ELECTROLITE [TRADE] corporation. Theassembled layers were then cured for an additional 30 minutes. Reactionwas catalyzed by a 0.5% vol/vol mixture of Irgacure 500 manufactured byCIBA-GIEGY Chemicals. The resulting elastomeric material exhibitedmoderate elasticity and adhered to glass.

Alternatively, other bonding methods may be used, including activatingthe elastomer surface, for example by plasma exposure, so that theelastomer layers/substrate will bond when placed in contact. Forexample, one possible approach to bonding together elastomer layerscomposed of the same material is set forth by Duffy et al, “RapidPrototyping of Microfluidic Systems in Poly (dimethylsiloxane),”Analytical Chemistry (1998), 70, 4974-4984, incorporated herein byreference. This paper discusses that exposing polydimethylsiloxane(PDMS) layers to oxygen plasma causes oxidation of the surface, withirreversible bonding occurring when the two oxidized layers are placedinto contact.

Yet another approach to bonding together successive layers of elastomeris to utilize the adhesive properties of uncured elastomer.Specifically, a thin layer of uncured elastomer such as RTV 615 [TRADE]is applied on top of a first cured elastomeric layer. Next, a secondcured elastomeric layer is placed on top of the uncured elastomericlayer. The thin middle layer of uncured elastomer is then cured toproduce a monolithic elastomeric structure. Alternatively, uncuredelastomer can be applied to the bottom of a first cured elastomer layer,with the first cured elastomer layer placed on top of a second curedelastomer layer. Curing the middle thin elastomer layer again results information of a monolithic elastomeric structure.

Where encapsulation of sacrificial layers is employed to fabricate theelastomer structure as described above in FIGS. 8-18, bonding ofsuccessive elastomeric layers may be accomplished by pouring uncuredelastomer over a previously cured elastomeric layer and any sacrificialmaterial patterned thereupon. Bonding between elastomer layers occursdue to interpenetration and reaction of the polymer chains of an uncuredelastomer layer with the polymer chains of a cured elastomer layer.Subsequent curing of the elastomeric layer will create a bond betweenthe elastomeric layers and create a monolithic elastomeric structure.

Referring to the first method of FIGS. 1 to 7B, first elastomeric layer20 may be created by spin-coating an RTV mixture on microfabricated mold12 at 2000 rpm's for 30 seconds yielding a thickness of approximately 40microns. Second elastomeric layer 22 may be created by spin-coating anRTV mixture on microfabricated mold 11. Both layers 20 and 22 may beseparately baked or cured at about 80° C. for 1.5 hours. The secondelastomeric layer 22 may be bonded onto first elastomeric layer 20 atabout 80° C. for about 1.5 hours.

Micromachined molds 10 and 12 may be patterned photoresist on siliconwafers. In an exemplary aspect, a SHIPLEY [®]-SJR 5740 [trade]photoresist was spun at 2000 rpm patterned with a high resolutiontransparency film as a mask and then developed yielding an inversechannel of approximately 10 microns in height. When baked atapproximately 200° C. for about 30 minutes, the photoresist reflows andthe inverse channels become rounded. In preferred aspects, the molds maybe treated with trimethylchlorosilane (TMCS) vapor for about a minutebefore each use in order to prevent adhesion of silicone rubber.

Using the various multilayer soft lithography construction techniquesand materials set forth herein, the present inventors haveexperimentally succeeded in creating channel networks comprises of up toseven separate elastomeric layers thick, with each layer being about 40μm thick. It is foreseeable that devices comprising more than sevenseparate elastomeric layers bonded together could be developed.

Suitable Elastomeric Materials:

Allcock et al, Contemporary Polymer Chemistry, 2^(nd) Ed. describeselastomers in general as polymers existing at a temperature betweentheir glass transition temperature and liquefaction temperature.Elastomeric materials exhibit elastic properties because the polymerchains readily undergo torsional motion to permit uncoiling of thebackbone chains in response to a force, with the backbone chainsrecoiling to assume the prior shape in the absence of the force. Ingeneral, elastomers deform when force is applied, but then return totheir original shape when the force is removed. The elasticity exhibitedby elastomeric materials may be characterized by a Young's modulus.Elastomeric materials having a Young's modulus of between about 1 Pa-1TPa, more preferably between about 10 Pa-100 GPa, more preferablybetween about 20 Pa-1 GPa, more preferably between about 50 Pa-10 MPa,and more preferably between about 100 Pa-1 MPa are useful in accordancewith the present invention, although elastomeric materials having aYoung's modulus outside of these ranges could also be utilized dependingupon the needs of a particular application.

The systems of the present invention may be fabricated from a widevariety of elastomers. In an exemplary aspect, elastomeric layers 20,22, 42, 46 and 50 may preferably be fabricated from silicone rubber.However, other suitable elastomers may also be used.

In an exemplary aspect of the present invention, the present systems arefabricated from an elastomeric polymer such as GE [®] RTV 15615 [trade](formulation), a vinyl-silane crosslinked (type) silicone elastomer(family). However, the present systems are not limited to this oneformulation, type or even this family of polymer; rather, nearly anyelastomeric polymer is suitable. An important requirement for thepreferred method of fabrication of the present microvalves is theability to bond multiple layers of elastomers together. In the case ofmultilayer soft lithography, layers of elastomer are cured separatelyand then bonded together. This scheme requires that cured layers possesssufficient reactivity to bond together. Either the layers may be of thesame type, and are capable of bonding to themselves, or they may be oftwo different types, and are capable of bonding to each other. Otherpossibilities include the use an adhesive between layers and the use ofthermoset elastomers.

Given the tremendous diversity of polymer chemistries, precursors,synthetic methods, reaction conditions, and potential additives, thereare a huge number of possible elastomer systems that could be used tomake monolithic elastomeric microvalves and pumps. Variations in thematerials used will most likely be driven by the need for particularmaterial properties, i.e. solvent resistance, stiffness, gaspermeability, or temperature stability.

There are many, many types of elastomeric polymers. A brief descriptionof the most common classes of elastomers is presented here, with theintent of showing that even with relatively “standard” polymers, manypossibilities for bonding exist. Common elastomeric polymers includepolyisoprene, polybutadiene, polychloroprene, polyisobutylene,poly(styrene-butadiene-styrene), the polyurethanes, and silicones.

Polyisoprene, Polybutadiene, Polychloroprene:

-   -   Polyisoprene, polybutadiene, and polychloroprene are all        polymerized from diene monomers, and therefore have one double        bond per monomer when polymerized. This double bond allows the        polymers to be converted to elastomers by vulcanization        (essentially, sulfur is used to form crosslinks between the        double bonds by heating). This would easily allow homogeneous        multilayer soft lithography by incomplete vulcanization of the        layers to be bonded; photoresist encapsulation would be possible        by a similar mechanism.

Polyisobutylene:

-   -   Pure polyisobutylene has no double bonds, but is crosslinked to        use as an elastomer by including a small amount (˜1%) of        isoprene in the polymerization. The isoprene monomers give        pendant double bonds on the polyisobutylene backbone, which may        then be vulcanized as above.

Poly(styrene-butadiene-styrene):

-   -   Poly(styrene-butadiene-styrene) is produced by living anionic        polymerization (that is, there is no natural chain-terminating        step in the reaction), so “live” polymer ends can exist in the        cured polymer. This makes it a natural candidate for the present        photoresist encapsulation system (where there will be plenty of        unreacted monomer in the liquid layer poured on top of the cured        layer). Incomplete curing would allow homogeneous multilayer        soft lithography (A to A bonding). The chemistry also        facilitates making one layer with extra butadiene (“A”) and        coupling agent and the other layer (“B”) with a butadiene        deficit (for heterogeneous multilayer soft lithography). SBS is        a “thermoset elastomer,” meaning that above a certain        temperature it melts and becomes plastic (as opposed to        elastic); reducing the temperature yields the elastomer again.        Thus, layers can be bonded together by heating.

Polyurethanes:

-   -   Polyurethanes are produced from di-isocyanates (A-A) and        di-alcohols or di-amines (B-B); since there are a large variety        of di-isocyanates and di-alcohols/amines, the number of        different types of polyurethanes is huge. The A vs. B nature of        the polymers, however, would make them useful for heterogeneous        multilayer soft lithography just as RTV 615 [trade] is: by using        excess A-A in one layer and excess B-B in the other layer.

Silicones:

-   -   Silicone polymers probably have the greatest structural variety,        and almost certainly have the greatest number of commercially        available formulations. The vinyl-to-(Si—H) crosslinking of RTV        615 [trade] (which allows both heterogeneous multilayer soft        lithography and photoresist encapsulation) has already been        discussed, but this is only one of several crosslinking methods        used in silicone polymer chemistry.

Cross Linking Agents:

In addition to the use of the simple “pure” polymers discussed above,crosslinking agents may be added. Some agents (like the monomers bearingpendant double bonds for vulcanization) are suitable for allowinghomogeneous (A to A) multilayer soft lithography or photoresistencapsulation; in such an approach the same agent is incorporated intoboth elastomer layers. Complementary agents (i.e. one monomer bearing apendant double bond, and another bearing a pendant Si—H group) aresuitable for heterogeneous (A to B) multilayer soft lithography. In thisapproach complementary agents are added to adjacent layers.

Other Materials:

In addition, polymers incorporating materials such as chlorosilanes ormethyl-, ethyl-, and phenylsilanes, and polydimethylsiloxane (PDMS) suchas DOW CHEMICAL [®] Corp. SYLGARD [®] 182, 184 or 186, or aliphaticurethane diacrylates such as (but not limited to) EbecrylEBECRYL [®] 270or IRR245 from UCB CHEMICAL [trade] may also be used.

The following is a non-exclusive list of elastomeric materials which maybe utilized in connection with the present invention: polyisoprene,polybutadiene, polychloroprene, polyisobutylene,poly(styrene-butadiene-styrene), the polyurethanes, and siliconepolymers; or poly(bis(fluoroalkoxy)phosphazene) (PNF, Eypel-F),poly(carborane-siloxanes) (Dexsil), poly(acrylonitrile-butadiene)(nitrile rubber), poly(1-butene),poly(chlorotrifluoroethylene-vinylidene fluoride) copolymers (Kel-F),poly(ethyl vinyl ether), poly(vinylidene fluoride), poly(vinylidenefluoride-hexafluoropropylene) copolymer (Viton), elastomericcompositions of polyvinylchloride (PVC), polysulfone, polycarbonate,polymethylmethacrylate (PMMA), and polytertrafluoroethylene (Teflon).

Doping and Dilution:

Elastomers may also be “doped” with uncrosslinkable polymer chains ofthe same class. For instance RTV 615 [trade] may be diluted with GE [®]SF96-50 [®] silicone fluid. This serves to reduce the viscosity of theuncured elastomer and reduces the Young's modulus of the curedelastomer. Essentially, the crosslink-capable polymer chains are spreadfurther apart by the addition of “inert” polymer chains, so this iscalled “dilution.” RTV 615 [trade] cures at up to 90% dilution, with adramatic reduction in Young's modulus.

FIG. 49 plots Young's modulus versus percentage dilution with GE SF96-50diluent of GE RTV 615 [trade] elastomer having a ratio of 30:1 A:B. FIG.49 shows that the flexibility of the elastomer material, and hence theresponsiveness of the valve membrane to an applied actuation force, canbe controlled during fabrication of the device.

Other examples of doping of elastomer material may include theintroduction of electrically conducting or magnetic species, asdescribed in detail below in conjunction with alternative methods ofactuating the membrane of the device. Should it be desired, doping withfine particles of material having an index of refraction different thanthe elastomeric material (i.e. silica, diamond, sapphire) is alsocontemplated as a system for altering the refractive index of thematerial. Strongly absorbing or opaque particles may be added to renderthe elastomer colored or opaque to incident radiation. This mayconceivably be beneficial in an optically addressable system.

Finally, by doping the elastomer with specific chemical species, thesedoped chemical species may be presented at the elastomer surface, thusserving as anchors or starting points for further chemicalderivitization.

Pre-Treatment and Surface Coating

Once the elastomeric material has been molded or etched into theappropriate shape, it may be necessary to pre-treat the material inorder to facilitate operation in connection with a particularapplication.

For example, one possible application for an elastomeric device inaccordance with the present invention is to sort biological entitiessuch as cells or DNA. In such an application, the hydrophobic nature ofthe biological entity may cause it to adhere to the hydrophobicelastomer of the walls of the channel. Therefore, it may be useful topre-treat the elastomeric structure order to impart a hydrophiliccharacter to the channel walls. In an embodiment of the presentinvention utilizing the GENERAL ELECTRIC RTV 615 [trade] elastomer, thiscan be accomplished by boiling the shaped elastomer in acid (e.g. 0.01%HCl in water, pH 2.7, at 60° C. for 40 min).

Other types of pre-treatment of elastomer material are also contemplatedby the present application. For example, certain portions of elastomermay be pre-treated to create anchors for surface chemistry reactions(for example in the formation of peptide chains), or binding sites forantibodies, as would be advantageous in a given application. Otherexamples of pre-treatment of elastomer material may include theintroduction of reflective material on the elastomer surface, asdescribed in detail below in conjunction with the micro-mirror arrayapplication.

Methods of Operating the Present Invention:

FIGS. 7B and 7H together show the closing of a first flow channel bypressurizing a second flow channel, with FIG. 7B (a front sectional viewcutting through flow channel 32 in corresponding FIG. 7A), showing anopen first flow channel 30; with FIG. 7H showing first flow channel 30closed by pressurization of the second flow channel 32.

Referring to FIG. 7B, first flow channel 30 and second flow channel 32are shown. Membrane 25 separates the flow channels, forming the top offirst flow channel 30 and the bottom of second flow channel 32. As canbe seen, flow channel 30 is “open.”

As can be seen in FIG. 7H, pressurization of flow channel 32 (either bygas or liquid introduced therein) causes membrane 25 to deflectdownward, thereby pinching off flow F passing through flow channel 30.Accordingly, by varying the pressure in channel 32, a linearly actuablevalving system is provided such that flow channel 30 can be opened orclosed by moving membrane 25 as desired. (For illustration purposesonly, channel 30 in FIG. 7G is shown in a “mostly closed” position,rather than a “fully closed” position).

It is to be understood that exactly the same valve opening and closingmethods can be achieved with flow channels 60 and 62.

Since such valves are actuated by moving the roof of the channelsthemselves (i.e.: moving membrane 25) valves and pumps produced by thistechnique have a truly zero dead volume, and switching valves made bythis technique have a dead volume approximately equal to the activevolume of the valve, for example about 100×100×10 μm=100 pL. Such deadvolumes and areas consumed by the moving membrane are approximately twoorders of magnitude smaller than known conventional microvalves. Smallerand larger valves and switching valves are contemplated in the presentinvention, and a non-exclusive list of ranges of dead volume includes 1aL to 1 uL, 100 aL to 100 nL, 1 fL to 10 nL, 100 fL to 1 nL, and 1 pL to100 pL.

The extremely small volumes capable of being delivered by pumps andvalves in accordance with the present invention represent a substantialadvantage. Specifically, the smallest known volumes of fluid capable ofbeing manually metered is around 0.1 μl. The smallest known volumescapable of being metered by automated systems is about ten-times larger(1 μl). Utilizing pumps and valves in accordance with the presentinvention, volumes of liquid of 10 nl or smaller can routinely bemetered and dispensed. The accurate metering of extremely small volumesof fluid enabled by the present invention would be extremely valuable ina large number of biological applications, including diagnostic testsand assays.

Equation 1 represents a highly simplified mathematical model ofdeflection of a rectangular, linear, elastic, isotropic plate of uniformthickness by an applied pressure:

w=(BPb ⁴)/(Eh ³), where:  (1)

-   -   w=deflection of plate;    -   B=shape coefficient (dependent upon length vs. width and support        of edges of plate);    -   P=applied pressure;    -   b=plate width    -   E=Young's modulus; and    -   h=plate thickness        Thus even in this extremely simplified expression, deflection of        an elastomeric membrane in response to a pressure will be a        function of: the length, width, and thickness of the membrane,        the flexibility of the membrane (Young's modulus), and the        applied actuation force. Because each of these parameters will        vary widely depending upon the actual dimensions and physical        composition of a particular elastomeric device in accordance        with the present invention, a wide range of membrane thicknesses        and elasticities, channel widths, and actuation forces are        contemplated by the present invention.

It should be understood that the formula just presented is only anapproximation, since in general the membrane does not have uniformthickness, the membrane thickness is not necessarily small compared tothe length and width, and the deflection is not necessarily smallcompared to length, width, or thickness of the membrane. Nevertheless,the equation serves as a useful guide for adjusting variable parametersto achieve a desired response of deflection versus applied force.

FIGS. 21 a and 21 b illustrate valve opening vs. applied pressure for a100 μm wide first flow channel 30 and a 50 μm wide second flow channel32. The membrane of this device was formed by a layer of GENERALELECTRIC [®] SILICONES RTV 615 [trade] having a thickness ofapproximately 30 μm and a Young's modulus of approximately 750 kPa.FIGS. 21 a and 21 b show the extent of opening of the valve to besubstantially linear over most of the range of applied pressures. Whilethe present invention does not require this linear actuation behavior,embodiments of the invention.

Air pressure was applied to actuate the membrane of the device through a10 cm long piece of plastic tubing having an outer diameter of 0.025″connected to a 25 mm piece of stainless steel hypodermic tubing with anouter diameter of 0.025″ and an inner diameter of 0.013.″ This tubingwas placed into contact with the control channel by insertion into theelastomeric block in a direction normal to the control channel. Airpressure was applied to the hypodermic tubing from an external LHDAminiature solenoid valve manufactured by Lee Co.

Connection of conventional microfluidic devices to an external fluidflow poses a number of problems avoided by the external configurationjust described. One such problem is the fragility of their connectionswith the external environment. Specifically, conventional microfluidicdevices are composed of hard, inflexible materials (such as silicon), towhich pipes or tubing allowing connection to external elements must bejoined. The rigidity of the conventional material creates significantphysical stress at points of contact with small and delicate externaltubing, rendering conventional microfluidic devices prone to fractureand leakage at these contact points.

By contrast, the elastomer of the present invention is flexible and canbe easily penetrated for external connection by a tube composed a hardmaterial. For example, in an elastomer structure fabricated utilizingthe method shown in FIGS. 1-7B, a hole extending from the exteriorsurface of the structure into the control channel can be made bypenetrating the elastomer with metal hypodermic tubing after the upperelastomer piece has been removed from the mold (as shown in FIG. 3) andbefore this piece has been bonded to the lower elastomer piece (as shownin FIG. 4). Between these steps, the roof of the control channel isexposed to the user's view and is accessible to insertion and properpositioning of the hole. Following completion of fabrication of thedevice, the metal hypodermic tubing is inserted into the hole tocomplete the fluid connection.

Moreover, the elastomer of the present invention will flex in responseto physical strain at the point of contact with an external connection,rendering the external physical connection more robust. This flexibilitysubstantially reduces the chance of leakage or fracture of the presentdevice.

Another disadvantage of conventional microfluidic devices is thedifficulty in establishing an effective seal between the device and itsexternal links. Because of the extremely narrow diameter of the channelsof these devices, even moderate rates of fluid flow can requireextremely high pressures. Unwanted leakage at the junction between thedevice and external connections may result. However, the flexibility ofthe elastomer of the present device also aids in overcoming leakagerelating to pressure. In particular, the flexible elastomeric materialflexes to conform around inserted tubing in order to form a pressureresistant seal.

While control of the flow of material through the device has so far beendescribed utilizing applied gas pressure, other fluids could be used.For example, air is compressible, and thus experiences some finite delaybetween the time of application of pressure by the external solenoidvalve and the time that this pressure is experienced by the membrane. Inan alternative embodiment of the present invention, pressure could beapplied from an external source to a noncompressible fluid such as wateror hydraulic oils, resulting in a near-instantaneous transfer of appliedpressure to the membrane. However, if the displaced volume of the valveis large or the control channel is narrow, higher viscosity of a controlfluid may contribute to delay in actuation. The optimal medium fortransferring pressure will therefore depend upon the particularapplication and device configuration, and both gaseous and liquid mediaare contemplated by the invention.

While external applied pressure as described above has been applied by apump/tank system through a pressure regulator and external miniaturevalve, other methods of applying external pressure are also contemplatedin the present invention, including gas tanks, compressors, pistonsystems, and columns of liquid. Also contemplated is the use ofnaturally occurring pressure sources such as may be found inside livingorganisms, such as blood pressure, gastric pressure, the pressurepresent in the cerebro-spinal fluid, pressure present in theintra-ocular space, and the pressure exerted by muscles during normalflexure. Other methods of regulating external pressure are alsocontemplated, such as miniature valves, pumps, macroscopic peristalticpumps, pinch valves, and other types of fluid regulating equipment suchas is known in the art.

As can be seen, the response of valves in accordance with embodiments ofthe present invention have been experimentally shown to be almostperfectly linear over a large portion of its range of travel, withminimal hysteresis. Accordingly, the present valves are ideally suitedfor microfluidic metering and fluid control. The linearity of the valveresponse demonstrates that the individual valves are well modeled asHooke's Law springs. Furthermore, high pressures in the flow channel(i.e.: back pressure) can be countered simply by increasing theactuation pressure. Experimentally, the present inventors have achievedvalve closure at back pressures of 70 kPa, but higher pressures are alsocontemplated. The following is a nonexclusive list of pressure rangesencompassed by the present invention: 10 Pa-25 MPa; 100 Pa-10 MPa, 1kPa-11 MPa, 1 kPa-1 MPa, 1 kPa-300 kPa, 5 kPa-200 kPa, and 15 kPa-100kPa.

While valves and pumps do not require linear actuation to open andclose, linear response does allow valves to more easily be used asmetering devices. In one embodiment of the invention, the opening of thevalve is used to control flow rate by being partially actuated to aknown degree of closure. Linear valve actuation makes it easier todetermine the amount of actuation force required to close the valve to adesired degree of closure. Another benefit of linear actuation is thatthe force required for valve actuation may be easily determined from thepressure in the flow channel. If actuation is linear, increased pressurein the flow channel may be countered by adding the same pressure (forceper unit area) to the actuated portion of the valve.

Linearity of a valve depends on the structure, composition, and methodof actuation of the valve structure. Furthermore, whether linearity is adesirable characteristic in a valve depends on the application.Therefore, both linearly and non-linearly actuatable valves arecontemplated in the present invention, and the pressure ranges overwhich a valve is linearly actuatable will vary with the specificembodiment.

FIG. 22 illustrates time response (i.e.: closure of valve as a functionof time in response to a change in applied pressure) of a 100 μm×100μm×10 μm RTV microvalve with 10-cm-long air tubing connected from thechip to a pneumatic valve as described above.

Two periods of digital control signal, actual air pressure at the end ofthe tubing and valve opening are shown in FIG. 22. The pressure appliedon the control line is 100 kPa, which is substantially higher than the˜40 kPa required to close the valve. Thus, when closing, the valve ispushed closed with a pressure 60 kPa greater than required. Whenopening, however, the valve is driven back to its rest position only byits own spring force (≦40 kPa). Thus, τ_(close) is expected to besmaller than τ_(open). There is also a lag between the control signaland control pressure response, due to the limitations of the miniaturevalve used to control the pressure. Calling such lags t and the 1/e timeconstants τ, the values are: t_(open)=3.63 ms, τ_(open)=1.88 ms,t_(close)=2.15 ms, τ_(close)=0.51 ms. If 3τ each are allowed for openingand closing, the valve runs comfortably at 75 Hz when filled withaqueous solution.

If one used another actuation method which did not suffer from openingand closing lag, this valve would run at ˜375 Hz. Note also that thespring constant can be adjusted by changing the membrane thickness; thisallows optimization for either fast opening or fast closing. The springconstant could also be adjusted by changing the elasticity (Young'smodulus) of the membrane, as is possible by introducing dopant into themembrane or by utilizing a different elastomeric material to serve asthe membrane (described above in conjunction with FIGS. 7C-7H.)

When experimentally measuring the valve properties as illustrated inFIGS. 21 and 22, the valve opening was measured by fluorescence. Inthese experiments, the flow channel was filled with a solution offluorescein isothiocyanate (FITC) in buffer (pH 8) and the fluorescenceof a square area occupying the center ˜⅓rd of the channel is monitoredon an epi-fluorescence microscope with a photomultiplier tube with a 10kHz bandwidth. The pressure was monitored with a Wheatstone-bridgepressure sensor (SENSYM [®] SCC15GD2 [trade]) pressurized simultaneouslywith the control line through nearly identical pneumatic connections.

Flow Channel Cross Sections:

The flow channels of the present invention may optionally be designedwith different cross sectional sizes and shapes, offering differentadvantages, depending upon their desired application. For example, thecross sectional shape of the lower flow channel may have a curved uppersurface, either along its entire length or in the region disposed underan upper cross channel). Such a curved upper surface facilitates valvesealing, as follows.

Referring to FIG. 19, a cross sectional view (similar to that of FIG.7B) through flow channels 30 and 32 is shown. As can be seen, flowchannel 30 is rectangular in cross sectional shape. In an alternatepreferred aspect of the invention, as shown in FIG. 20, thecross-section of a flow channel 30 instead has an upper curved surface.

Referring first to FIG. 19, when flow channel 32 is pressurized, themembrane portion 25 of elastomeric block 24 separating flow channels 30and 32 will move downwardly to the successive positions shown by thedotted lines 25A, 25B, 25C, 25D, and 25E. As can be seen, incompletesealing may possibly result at the edges of flow channel 30 adjacentplanar substrate 14.

In the alternate preferred embodiment of FIG. 20, flow channel 30 a hasa curved upper wall 25A. When flow channel 32 is pressurized, membraneportion 25 will move downwardly to the successive positions shown bydotted lines 25A2, 25A3, 25A4 and 25A5, with edge portions of themembrane moving first into the flow channel, followed by top membraneportions. An advantage of having such a curved upper surface at membrane25A is that a more complete seal will be provided when flow channel 32is pressurized. Specifically, the upper wall of the flow channel 30 willprovide a continuous contacting edge against planar substrate 14,thereby avoiding the “island” of contact seen between wall 25 and thebottom of flow channel 30 in FIG. 19.

Another advantage of having a curved upper flow channel surface atmembrane 25A is that the membrane can more readily conform to the shapeand volume of the flow channel in response to actuation. Specifically,where a rectangular flow channel is employed, the entire perimeter (2×flow channel height, plus the flow channel width) must be forced intothe flow channel. However where an arched flow channel is used, asmaller perimeter of material (only the semi-circular arched portion)must be forced into the channel. In this manner, the membrane requiresless change in perimeter for actuation and is therefore more responsiveto an applied actuation force to block the flow channel

In an alternate aspect, (not illustrated), the bottom of flow channel 30is rounded such that its curved surface mates with the curved upper wall25A as seen in FIG. 20 described above.

In summary, the actual conformational change experienced by the membraneupon actuation will depend upon the configuration of the particularelastomeric structure. Specifically, the conformational change willdepend upon the length, width, and thickness profile of the membrane,its attachment to the remainder of the structure, and the height, width,and shape of the flow and control channels and the material propertiesof the elastomer used. The conformational change may also depend uponthe method of actuation, as actuation of the membrane in response to anapplied pressure will vary somewhat from actuation in response to amagnetic or electrostatic force.

Moreover, the desired conformational change in the membrane will alsovary depending upon the particular application for the elastomericstructure. In the simplest embodiments described above, the valve mayeither be open or closed, with metering to control the degree of closureof the valve. In other embodiments however, it may be desirable to alterthe shape of the membrane and/or the flow channel in order to achievemore complex flow regulation. For instance, the flow channel could beprovided with raised protrusions beneath the membrane portion, such thatupon actuation the membrane shuts off only a percentage of the flowthrough the flow channel, with the percentage of flow blockedinsensitive to the applied actuation force.

Many membrane thickness profiles and flow channel cross-sections arecontemplated by the present invention, including rectangular,trapezoidal, circular, ellipsoidal, parabolic, hyperbolic, andpolygonal, as well as sections of the above shapes. More complexcross-sectional shapes, such as the embodiment with protrusionsdiscussed immediately above or an embodiment having concavities in theflow channel, are also contemplated by the present invention.

In addition, while the invention is described primarily above inconjunction with an embodiment wherein the walls and ceiling of the flowchannel are formed from elastomer, and the floor of the channel isformed from an underlying substrate, the present invention is notlimited to this particular orientation. Walls and floors of channelscould also be formed in the underlying substrate, with only the ceilingof the flow channel constructed from elastomer. This elastomer flowchannel ceiling would project downward into the channel in response toan applied actuation force, thereby controlling the flow of materialthrough the flow channel. In general, monolithic elastomer structures asdescribed elsewhere in the instant application are preferred formicrofluidic applications. However, it may be useful to employ channelsformed in the substrate where such an arrangement provides advantages.For instance, a substrate including optical waveguides could beconstructed so that the optical waveguides direct light specifically tothe side of a microfluidic channel.

Alternate Valve Actuation Techniques:

In addition to pressure based actuation systems described above,optional electrostatic and magnetic actuation systems are alsocontemplated, as follows.

Electrostatic actuation can be accomplished by forming oppositelycharged electrodes (which will tend to attract one another when avoltage differential is applied to them) directly into the monolithicelastomeric structure. For example, referring to FIG. 7B, an optionalfirst electrode 70 (shown in phantom) can be positioned on (or in)membrane 25 and an optional second electrode 72 (also shown in phantom)can be positioned on (or in) planar substrate 14. When electrodes 70 and72 are charged with opposite polarities, an attractive force between thetwo electrodes will cause membrane 25 to deflect downwardly, therebyclosing the “valve” (i.e.: closing flow channel 30).

For the membrane electrode to be sufficiently conductive to supportelectrostatic actuation, but not so mechanically stiff so as to impedethe valve's motion, a sufficiently flexible electrode must be providedin or over membrane 25. Such an electrode may be provided by a thinmetallization layer, doping the polymer with conductive material, ormaking the surface layer out of a conductive material.

In an exemplary aspect, the electrode present at the deflecting membranecan be provided by a thin metallization layer which can be provided, forexample, by sputtering a thin layer of metal such as 20 nm of gold. Inaddition to the formation of a metallized membrane by sputtering, othermetallization approaches such as chemical epitaxy, evaporation,electroplating, and electroless plating are also available. Physicaltransfer of a metal layer to the surface of the elastomer is alsoavailable, for example by evaporating a metal onto a flat substrate towhich it adheres poorly, and then placing the elastomer onto the metaland peeling the metal off of the substrate.

A conductive electrode 70 may also be formed by depositing carbon black(i.e. CABOT VULCAN XC72R) [trade] on the elastomer surface, either bywiping on the dry powder or by exposing the elastomer to a suspension ofcarbon black in a solvent which causes swelling of the elastomer, (suchas a chlorinated solvent in the case of PDMS). Alternatively, theelectrode 70 may be formed by constructing the entire layer 20 out ofelastomer doped with conductive material (i.e. carbon black or finelydivided metal particles). Yet further alternatively, the electrode maybe formed by electrostatic deposition, or by a chemical reaction thatproduces carbon. In experiments conducted by the present inventors,conductivity was shown to increase with carbon black concentration from5.6×10⁻¹⁶ to about 5×10⁻³ (Ω-cm)⁻¹. The lower electrode 72, which is notrequired to move, may be either a compliant electrode as describedabove, or a conventional electrode such as evaporated gold, a metalplate, or a doped semiconductor electrode.

Alternatively, magnetic actuation of the flow channels can be achievedby fabricating the membrane separating the flow channels with amagnetically polarizable material such as iron, or a permanentlymagnetized material such as polarized NdFeB. In experiments conducted bythe present inventors, magnetic silicone was created by the addition ofiron powder (about 1 um particle size), up to 20% iron by weight.

Where the membrane is fabricated with a magnetically polarizablematerial, the membrane can be actuated by attraction in response to anapplied magnetic field. Where the membrane is fabricated with a materialcapable of maintaining permanent magnetization, the material can firstbe magnetized by exposure to a sufficiently high magnetic field, andthen actuated either by attraction or repulsion in response to thepolarity of an applied inhomogeneous magnetic field.

The magnetic field causing actuation of the membrane can be generated ina variety of ways. In one embodiment, the magnetic field is generated byan extremely small inductive coil formed in or proximate to theelastomer membrane. The actuation effect of such a magnetic coil wouldbe localized, allowing actuation of individual pump and/or valvestructures. Alternatively, the magnetic field could be generated by alarger, more powerful source, in which case actuation would be globaland would actuate multiple pump and/or valve structures at one time.

It is further possible to combine pressure actuation with electrostaticor magnetic actuation. Specifically, a bellows structure in fluidcommunication with a recess could be electrostatically or magneticallyactuated to change the pressure in the recess and thereby actuate amembrane structure adjacent to the recess.

In addition to electrical or magnetic actuation as described above,optional electrolytic and electrokinetic actuation systems are alsocontemplated by the present invention. For example, actuation pressureon the membrane could arise from an electrolytic reaction in a recessoverlying the membrane. In such an embodiment, electrodes present in therecess would apply a voltage across an electrolyte in the recess. Thispotential difference would cause electrochemical reaction at theelectrodes and result in the generation of gas species, giving rise to apressure differential in the recess.

Alternatively, actuation pressure on the membrane could arise from anelectrokinetic fluid flow in the control channel. In such an embodiment,electrodes present at opposite ends of the control channel would apply apotential difference across an electrolyte present in the controlchannel. Migration of charged species in the electrolyte to therespective electrodes could give rise to a pressure differential.

Finally, it is also possible to actuate the device by causing a fluidflow in the control channel based upon the application of thermalenergy, either by thermal expansion or by production of gas from liquid.Similarly, chemical reactions generating gaseous products may produce anincrease in pressure sufficient for membrane actuation.

Networked Systems:

FIGS. 23A and 23B show a views of a single on/off valve, identical tothe systems set forth above, (for example in FIG. 7A). FIGS. 24A and 24Bshows a peristaltic pumping system comprised of a plurality of thesingle addressable on/off valves as seen in FIG. 23, but networkedtogether. FIG. 25 is a graph showing experimentally achieved pumpingrates vs. frequency for the peristaltic pumping system of FIG. 24. FIGS.26A and 26B show a schematic view of a plurality of flow channels whichare controllable by a single control line. This system is also comprisedof a plurality of the single addressable on/off valves of FIG. 23,multiplexed together, but in a different arrangement than that of FIG.23. FIG. 27 is a schematic illustration of a multiplexing system adaptedto permit fluid flow through selected channels, comprised of a pluralityof the single on/off valves of FIG. 23, joined or networked together.

Referring first to FIGS. 23A and 23B, a schematic of flow channels 30and 32 is shown. Flow channel 30 preferably has a fluid (or gas) flow Fpassing therethrough.

Flow channel 32, (which crosses over flow channel 30, as was alreadyexplained herein), is pressurized such that membrane 25 separating theflow channels may be depressed into the path of flow channel 30,shutting off the passage of flow F therethrough, —as has been explained.As such, “flow channel” 32 can also be referred to as a “control line”which actuates a single valve in flow channel 30. In FIGS. 23 to 26, aplurality of such addressable valves are joined or networked together invarious arrangements to produce pumps, capable of peristaltic pumping,and other fluidic logic applications.

Referring to FIGS. 24A and 24B, a system for peristaltic pumping isprovided, as follows. A flow channel 30 has a plurality of generallyparallel flow channels (i.e.: control lines) 32A, 32B and 32C passingthereover. By pressurizing control line 32A, flow F through flow channel30 is shut off under membrane 25A at the intersection of control line32A and flow channel 30. Similarly, (but not shown), by pressurizingcontrol line 32B, flow F through flow channel 30 is shut off undermembrane 25B at the intersection of control line 32B and flow channel30, etc.

Each of control lines 32A, 32B, and 32C is separately addressable.Therefore, peristalsis may be actuated by the pattern of actuating 32Aand 32C together, followed by 32A, followed by 32A and 32B together,followed by 32B, followed by 32B and C together, etc. This correspondsto a successive “101, 100, 110, 010, 011, 001” pattern, where “0”indicates “valve open” and “1” indicates “valve closed.” Thisperistaltic pattern is also known as a 120° pattern (referring to thephase angle of actuation between three valves). Other peristalticpatterns are equally possible, including 60° and 90° patterns.

In experiments performed by the inventors, a pumping rate of 2.35 nL/swas measured by measuring the distance traveled by a column of water inthin (0.5 mm i.d.) tubing; with 100×100×10 μm valves under an actuationpressure of 40 kPa. The pumping rate increased with actuation frequencyuntil approximately 75 Hz, and then was nearly constant until above 200Hz. The valves and pumps are also quite durable and the elastomermembrane, control channels, or bond have never been observed to fail. Inexperiments performed by the inventors, none of the valves in theperistaltic pump described herein show any sign of wear or fatigue aftermore than 4 million actuations. In addition to their durability, theyare also gentle. A solution of E. coli pumped through a channel andtested for viability showed a 94% survival rate.

FIG. 25 is a graph showing experimentally achieved pumping rates vs.frequency for the peristaltic pumping system of FIG. 24.

FIGS. 26A and 26B illustrates another way of assembling a plurality ofthe addressable valves of FIG. 21. Specifically, a plurality of parallelflow channels 30A, 30B, and 30C are provided. Flow channel (i.e.:control line) 32 passes thereover across flow channels 30A, 30B, and30C. Pressurization of control line 32 simultaneously shuts off flowsF1, F2 and F3 by depressing membranes 25A, 25B, and 25C located at theintersections of control line 32 and flow channels 30A, 30B, and 30C.

FIG. 27 is a schematic illustration of a multiplexing system adapted toselectively permit fluid to flow through selected channels, as follows.The downward deflection of membranes separating the respective flowchannels from a control line passing thereabove (for example, membranes25A, 25B, and 25C in FIGS. 26A and 26B) depends strongly upon themembrane dimensions. Accordingly, by varying the widths of flow channelcontrol line 32 in FIGS. 26A and 26B, it is possible to have a controlline pass over multiple flow channels, yet only actuate (i.e.: seal)desired flow channels. FIG. 27 illustrates a schematic of such a system,as follows.

A plurality of parallel flow channels 30A, 30B, 30C, 30D, 30E and 30Fare positioned under a plurality of parallel control lines 32A, 32B,32C, 32D, 32E and 32F. Control channels 32A, 32B, 32C, 32D, 32E and 32Fare adapted to shut off fluid flows F1, F2, F3, F4, F5 and F6 passingthrough parallel flow channels 30A, 30B, 30C, 30D, 30E and 30F using anyof the valving systems described above, with the following modification.

Each of control lines 32A, 32B, 32C, 32D, 32E and 32F have both wide andnarrow portions. For example, control line 32A is wide in locationsdisposed over flow channels 30A, 30C and 30E. Similarly, control line32B is wide in locations disposed over flow channels 30B, 30D and 30F,and control line 32C is wide in locations disposed over flow channels30A, 30B, 30E and 30F.

At the locations where the respective control line is wide, itspressurization will cause the membrane (25) separating the flow channeland the control line to depress significantly into the flow channel,thereby blocking the flow passage therethrough. Conversely, in thelocations where the respective control line is narrow, membrane (25)will also be narrow. Accordingly, the same degree of pressurization willnot result in membrane (25) becoming depressed into the flow channel(30). Therefore, fluid passage thereunder will not be blocked.

For example, when control line 32A is pressurized, it will block flowsF1, F3 and F5 in flow channels 30A, 30C and 30E. Similarly, when controlline 32C is pressurized, it will block flows F1, F2, F5 and F6 in flowchannels 30A, 30B, 30E and 30F. As can be appreciated, more than onecontrol line can be actuated at the same time. For example, controllines 32A and 32C can be pressurized simultaneously to block all fluidflow except F4 (with 32A blocking F1, F3 and F5; and 32C blocking F1,F2, F5 and F6).

By selectively pressurizing different control lines (32) both togetherand in various sequences, a great degree of fluid flow control can beachieved. Moreover, by extending the present system to more than sixparallel flow channels (30) and more than four parallel control lines(32), and by varying the positioning of the wide and narrow regions ofthe control lines, very complex fluid flow control systems may befabricated. A property of such systems is that it is possible to turn onany one flow channel out of n flow channels with only 2(log₂ n) controllines.

The inventors have succeeded in fabricating microfluidic structures withdensities of 30 devices/mm², but greater densities are achievable.

Selectively Addressable Reaction Chambers Along Flow Lines:

In a further embodiment of the invention, illustrated in FIGS. 28A, 28B,28C and 28D, a system for selectively directing fluid flow into one moreof a plurality of reaction chambers disposed along a flow line isprovided.

FIG. 28A shows a top view of a flow channel 30 having a plurality ofreaction chambers 80A and 80B disposed therealong. Preferably flowchannel 30 and reaction chambers 80A and 80B are formed together asrecesses into the bottom surface of a first layer 100 of elastomer.

FIG. 28B shows a bottom plan view of another elastomeric layer 110 withtwo control lines 32A and 32B each being generally narrow, but havingwide extending portions 33A and 33B formed as recesses therein.

As seen in the exploded view of FIG. 28C, and assembled view of FIG.28D, elastomeric layer 110 is placed over elastomeric layer 100. Layers100 and 110 are then bonded together, and the integrated system operatesto selectively direct fluid flow F (through flow channel 30) into eitheror both of reaction chambers 80A and 80B, as follows. Pressurization ofcontrol line 32A will cause the membrane 25 (i.e., the thin portion ofelastomer layer 100 located below extending portion 33A and over regions82A of reaction chamber 80A) to become depressed, thereby shutting offfluid flow passage in regions 82A, effectively sealing reaction chamber80 from flow channel 30. As can also be seen, extending portion 33A iswider than the remainder of control line 32A. As such, pressurization ofcontrol line 32A will not result in control line 32A sealing flowchannel 30.

As can be appreciated, either or both of control lines 32A and 32B canbe actuated at once. When both control lines 32A and 32B are pressurizedtogether, sample flow in flow channel 30 will enter neither of reactionchambers 80A or 80B.

The concept of selectably controlling fluid introduction into variousaddressable reaction chambers disposed along a flow line (FIG. 28) canbe combined with concept of selectably controlling fluid flow throughone or more of a plurality of parallel flow lines (FIG. 27) to yield asystem in which a fluid sample or samples can be can be sent to anyparticular reaction chamber in an array of reaction chambers. An exampleof such a system is provided in FIG. 29, in which parallel controlchannels 32A, 32B and 32C with extending portions 34 (all shown inphantom) selectively direct fluid flows F1 and F2 into any of the arrayof reaction wells 80A, 80B, 80C or 80D as explained above; whilepressurization of control lines 32C and 32D selectively shuts off flowsF2 and F1, respectively.

In yet another novel embodiment, fluid passage between parallel flowchannels is possible. Referring to FIG. 30, either or both of controllines 32A or 32D can be depressurized such that fluid flow throughlateral passageways 35 (between parallel flow channels 30A and 30B) ispermitted. In this aspect of the invention, pressurization of controllines 32C and 32D would shut flow channel 30A between 35A and 35B, andwould also shut lateral passageways 35B. As such, flow entering as flowF1 would sequentially travel through 30A, 35A and leave 30B as flow F4.

Switchable Flow Arrays

In yet another novel embodiment, fluid passage can be selectivelydirected to flow in either of two perpendicular directions. An exampleof such a “switchable flow array” system is provided in FIGS. 31A to31D. FIG. 31A shows a bottom view of a first layer of elastomer 90, (orany other suitable substrate), having a bottom surface with a pattern ofrecesses forming a flow channel grid defined by an array of solid posts92, each having flow channels passing therearound.

In preferred aspects, an additional layer of elastomer is bound to thetop surface of layer 90 such that fluid flow can be selectively directedto move either in direction F1, or perpendicular direction F2. FIG. 31is a bottom view of the bottom surface of the second layer of elastomer95 showing recesses formed in the shape of alternating “vertical”control lines 96 and “horizontal” control lines 94. “Vertical” controllines 96 have the same width therealong, whereas “horizontal” controllines 94 have alternating wide and narrow portions, as shown.

Elastomeric layer 95 is positioned over top of elastomeric layer 90 suchthat “vertical” control lines 96 are positioned over posts 92 as shownin FIG. 31C and “horizontal” control lines 94 are positioned with theirwide portions between posts 92, as shown in FIG. 31D.

As can be seen in FIG. 31C, when “vertical” control lines 96 arepressurized, the membrane of the integrated structure formed by theelastomeric layer initially positioned between layers 90 and 95 inregions 98 will be deflected downwardly over the array of flow channelssuch that flow in only able to pass in flow direction F2 (i.e.:vertically), as shown.

As can be seen in FIG. 31D, when “horizontal” control lines 94 arepressurized, the membrane of the integrated structure formed by theelastomeric layer initially positioned between layers 90 and 95 inregions 99 will be deflected downwardly over the array of flow channels,(but only in the regions where they are widest), such that flow in onlyable to pass in flow direction F1 (i.e.: horizontally), as shown.

The design illustrated in FIG. 31 allows a switchable flow array to beconstructed from only two elastomeric layers, with no vertical viaspassing between control lines in different elastomeric layers required.If all vertical flow control lines 94 are connected, they may bepressurized from one input. The same is true for all horizontal flowcontrol lines 96.

Biopolymer Synthesis

The present elastomeric valving structures can also be used inbiopolymer synthesis, for example, in synthesizing oligonucleotides,proteins, peptides, DNA, etc. In a preferred aspect, such biopolymersynthesis systems may comprise an integrated system comprising an arrayof reservoirs, fluidic logic (according to the present invention) forselecting flow from a particular reservoir, an array of channels orreservoirs in which synthesis is performed, and fluidic logic (alsoaccording to the present invention) for determining into which channelsthe selected reagent flows. An example of such a system 200 isillustrated in FIG. 32, as follows.

Four reservoirs 150A, 150B, 150C and 150D have bases A, C, T and Grespectively disposed therein, as shown. Four flow channels 30A, 30B,30C and 30D are connected to reservoirs 150A, 150B, 150C and 150D. Fourcontrol lines 32A, 32B, 32C and 32D (shown in-phantom) are disposedthereacross with control line 32A permitting flow only through flowchannel 30A (i.e.: sealing flow channels 30B, 30C and 30D), when controlline 32A is pressurized. Similarly, control line 32B permits flow onlythrough flow channel 30B when pressurized. As such, the selectivepressurization of control lines 32A, 32B, 32C and 32D sequentiallyselects a desired base A, C, T and G from a desired reservoir 150A,150B, 150C or 150D. The fluid then passes through flow channel 120 intoa multiplexed channel flow controller 125, (including, for example, anysystem as shown in FIGS. 26A to 31D) which in turn directs fluid flowinto one or more of a plurality of synthesis channels or chambers 122A,122B, 122C, 122D or 122E in which solid phase synthesis may be carriedout.

FIG. 33 shows a further extension of this system on which a plurality ofreservoirs R1 to R13 (which may contain bases A, T, C and G, or anyother reactants, such as would be used in combinatorial chemistry), areconnected to systems 200 as set forth in FIG. 32. Systems 200 areconnected to a multiplexed channel flow controller 125, (including, forexample, any system as shown in FIGS. 26A to 31D) which is in turnconnected to a switchable flow array (for example as shown in FIG. 31).An advantage of this system is that both of multiplexed channel flowcontrollers 125 and fluid selection systems 200 can be controlled by thesame pressure inputs 170 and 172, provided a “close horizontal” and a“close vertical” control lines (160 and 162, in phantom) are alsoprovided.

In further alternate aspects of the invention, a plurality ofmultiplexed channel flow controllers (such as 125)—may be used, witheach flow controller initially positioned stacked above one another on adifferent elastomeric layer, with vertical vias or interconnects betweenthe elastomer layers (which may be created by lithographicallypatterning an etch resistant layer on top of a elastomer layer, thenetching the elastomer and finally removing the etch resist before addingthe last layer of elastomer).

For example, a vertical via in an elastomer layer can be created byetching a hole down onto a raised line on a micromachined mold, andbonding the next layer such that a channel passes over that hole. Inthis aspect of the invention, multiple synthesis with a plurality ofmultiplexed channel flow controllers 125 is possible.

The bonding of successive layers of molded elastomer to form amulti-layer structure is shown in FIG. 34, which is an opticalmicrograph of a section of a test structure composed of seven layers ofelastomer. The scale bar of FIG. 34 is 200 μm.

One method for fabricating an elastomer layer having the vertical viafeature utilized in a multi-layer structure is shown in FIGS. 35A-35D.FIG. 35A shows formation of elastomer layer 3500 over micromachined mold3502 including raised line 3502 a.

FIG. 35B shows formation of metal etch blocking layer 3504 overelastomer layer 3500, followed by the patterning of photoresist mask3506 over etch blocking layer 3504 to cover masked regions 3508 andleave exposed unmasked regions 3510. FIG. 35C shows the exposure tosolvent which removes etch blocking layer 3504 in unmasked regions 3510.

FIG. 35D shows removal of the patterned photoresist, followed bysubsequent etching of underlying elastomer 3500 in unmasked regions 3510to form vertical via 3512. Subsequent exposure to solvent removesremaining etch blocking layer 3504 in masked regions 3508 selective tothe surrounding elastomer 3500 and mold 3502. This elastomer layer maythen be incorporated into an elastomer structure by multilayer softlithography.

This series of steps can be repeated as necessary to form amulti-layered structure having the desired number and orientation ofvertical vias between channels of successive elastomer layers.

The inventors of the present invention have succeeded in etching viasthrough GE [0]RTV 615 [trade] layers using a solution ofTetrabutylammonium fluoride in organic solvent. Gold serves as the etchblocking material, with gold removed selective to GE [®] RTV 615 [trade]utilizing a KI/I₂/H₂O mixture.

Alternatively, vertical vias between channels in successive elastomerlayers could be formed utilizing a negative mask technique. In thisapproach, a negative mask of a metal foil is patterned, and subsequentformation of an etch blocking layer is inhibited where the metal foil ispresent. Once the etch blocking material is patterned, the negativemetal foil mask is removed, permitting selective etching of theelastomer as described above.

In yet another approach, vertical vias could be formed in an elastomerlayer using ablation of elastomer material through application ofradiation from an applied laser beam.

While the above approach is described in connection with the synthesisof biopolymers, the invention is not limited to this application. Thepresent invention could also function in a wide variety of combinatorialchemical synthesis approaches.

Other Applications:

Advantageous applications of the present monolithic microfabricatedelastomeric valves and pumps are numerous. Accordingly, the presentinvention is not limited to any particular application or use thereof.In preferred aspects, the following uses and applications for thepresent invention are contemplated.

1. Cell/DNA Sorting

The present microfluidic pumps and valves can also be used in flowcytometers for cell sorting and DNA sizing. Sorting of objects basedupon size is extremely useful in many technical fields.

For example, many assays in biology require determination of the size ofmolecular-sized entities. Of particular importance is the measurement oflength distribution of DNA molecules in a heterogeneous solution. Thisis commonly done using gel electrophoresis, in which the molecules areseparated by their differing mobility in a gel matrix in an appliedelectric field, and their positions detected by absorption or emissionof radiation. The lengths of the DNA molecules are then inferred fromtheir mobility.

While powerful, electrophoretic methods pose disadvantages. For mediumto large DNA molecules, resolution, i.e. the minimum length differenceat which different molecular lengths may be distinguished, is limited toapproximately 10% of the total length. For extremely large DNAmolecules, the conventional sorting procedure is not workable. Moreover,gel electrophoresis is a relatively lengthy procedure, and may requireon the order of hours or days to perform.

The sorting of cellular-sized entities is also an important task.Conventional flow cell sorters are designed to have a flow chamber witha nozzle and are based on the principle of hydrodynamic focusing withsheath flow. Most conventional cell sorters combine the technology ofpiezo-electric drop generation and electrostatic deflection to achievedroplet generation and high sorting rates. However, this approach offerssome important disadvantages. One disadvantage is that the complexity,size, and expense of the sorting device requires that it be reusable inorder to be cost-effective. Reuse can in turn lead to problems withresidual materials causing contamination of samples and turbulent fluidflow.

Therefore, there is a need in the art for a simple, inexpensive, andeasily fabricated sorting device which relies upon the mechanicalcontrol of fluid flow rather than upon electrical interactions betweenthe particle and the solute.

FIG. 36 shows one embodiment of a sorting device in accordance with thepresent invention. Sorting device 3600 is formed from a switching valvestructure created from channels present in an elastomeric block.Specifically, flow channel 3602 is T-shaped, with stem 3602 a of flowchannel 3602 in fluid communication with sample reservoir 3604containing sortable entities 3606 of different types denoted by shape(square, circle, triangle, etc.). Left branch 3602 b of flow channel3602 is in fluid communication with waste reservoir 3608. Right branch3602 c of flow channel 3602 is in communication with collectionreservoir 3610.

Control channels 3612 a, 3612 b, and 3612 c overlie and are separatedfrom stem 3602 a of flow channel 3602 by elastomeric membrane portions3614 a, 3614 b, and 3614 c respectively. Together, stem 3602 a of flowchannel 3602 and control channels 3612 a, 3612 b, and 3612 c form firstperistaltic pump structure 3616 similar to that described at lengthabove in connection with FIG. 24 a.

Control channel 3612 d overlies and is separated from right branch 3602c of flow channel 3602 by elastomeric membrane portion 3614 d. Together,right branch 3602 c of flow channel 3602 and control channels 3612 dforms first valve structure 3618 a. Control channel 3612 e overlies andis separated from left branch 3602 c of flow channel 3602 by elastomericmembrane portion 3614 e. Together, left branch 3602 c of flow channel3602 and control channel 3612 e forms second valve structure 3618 b.

As shown in FIG. 36, stem 3602 a of flow channel 3602 narrowsconsiderably as it approaches detection window 3620 adjacent to thejunction of stem 3602 a, right branch 3602 b, and left branch 3602 c.Detection window 3620 is of sufficient width to allow for uniformillumination of this region. In one embodiment, the width of the stemnarrows from 100 μm to 5 μm at the detection window. The width of thestem at the detection window can be precisely formed using the softlithography or photoresist encapsulation fabrication techniquesdescribed extensively above, and will be depend upon the nature and sizeof the entity to be sorted.

Operation of sorting device in accordance with one embodiment of thepresent invention is as follows.

The sample is diluted to a level such that only a single sortable entitywould be expected to be present in the detection window at any time.Peristaltic pump 3616 is activated by flowing a fluid through controlchannels 3612 a-c as described extensively above. In addition, secondvalve structure 3618 b is closed by flowing fluid through controlchannel 3612 e. As a result of the pumping action of peristaltic pump3616 and the blocking action of second valve 3618 b, fluid flows fromsample reservoir 3604 through detection window 3620 into waste reservoir3608. Because of the narrowing of stem 3604, sortable entities presentin sample reservoir 3604 are carried by this regular fluid flow, one ata time, through detection window 3620.

Radiation 3640 from source 3642 is introduced into detection window3620. This is possible due to the transmissive property of theelastomeric material. Absorption or emission of radiation 3640 bysortable entity 3606 is then detected by detector 3644.

If sortable entity 3606 a within detection window 3620 is intended to besegregated and collected by sorting device 3600, first valve 3618 a isactivated and second valve 3618 b is deactivated. This has the effect ofdrawing sortable entity 3606 a into collection reservoir 3610, and atthe same time transferring second sortable entity 3606 b into detectionwindow 3620. If second sortable entity 3602 b is also identified forcollection, peristaltic pump 3616 continues to flow fluid through rightbranch 3602 c of flow channel 3602 into collection reservoir 3610.However, if second entity 3606 b is not to be collected, first valve3618 a opens and second valve 3618 b closes, and first peristaltic pump3616 resumes pumping liquid through left branch 3602 b of flow channel3602 into waste reservoir 3608.

While one specific embodiment of a sorting device and a method foroperation thereof is described in connection with FIG. 36, the presentinvention is not limited to this embodiment. For example, fluid need notbe flowed through the flow channels using the peristaltic pumpstructure, but could instead be flowed under pressure with theelastomeric valves merely controlling the directionality of flow. In yetanother embodiment, a plurality of sorting structures could be assembledin series in order to perform successive sorting operations, with thewaste reservoir of FIG. 36 simply replaced by the stem of the nextsorting structure.

Moreover, a high throughput method of sorting could be employed, whereina continuous flow of fluid from the sample reservoir through the windowand junction into the waste reservoir is maintained until an entityintended for collection is detected in the window. Upon detection of anentity to be collected, the direction of fluid flow by the pumpstructure is temporarily reversed in order to transport the desiredparticle back through the junction into the collection reservoir. Inthis manner, the sorting device could utilize a higher flow rate, withthe ability to backtrack when a desired entity is detected.

Such an alternative high throughput sorting technique could be used whenthe entity to be collected is rare, and the need to backtrackinfrequent.

Sorting in accordance with the present invention would avoid thedisadvantages of sorting utilizing conventional electrokinetic flow,such as bubble formation, a strong dependence of flow magnitude anddirection on the composition of the solution and surface chemistryeffects, a differential mobility of different chemical species, anddecreased viability of living organisms in the mobile medium.

2. Semiconductor Processing

Systems for semiconductor gas flow control, (particularly for epitaxialapplications in which small quantities of gases are accurately metered),are also contemplated by the present invention. For example, duringfabrication of semiconductor devices solid material is deposited on topof a semiconductor substrate utilizing chemical vapor deposition (CVD).This is accomplished by exposing the substrate to a mixture of gasprecursor materials, such that these gases react and the resultingproduct crystallizes on top of the substrate.

During such CVD processes, conditions must be carefully controlled toensure uniform deposition of material free of defects that could degradethe operation of the electrical device. One possible source ofnonuniformity is variation in the flow rate of reactant gases to theregion over the substrate. Poor control of the gas flow rate can alsolead to variations in the layer thicknesses from run to run, which isanother source of error. Unfortunately, there has been a significantproblem in controlling the amount of gas flowed into the processingchamber, and maintaining stable flow rates in conventional gas deliverysystems.

Accordingly, FIG. 37A shows one embodiment of the present inventionadapted to convey, at precisely-controllable flow rates, processing gasover the surface of a semiconductor wafer during a CVD process.Specifically, semiconductor wafer 3700 is positioned upon wafer support3702 located within a CVD chamber. Elastomeric structure 3704 containinga large number of evenly distributed orifices 3706 is positioned justabove the surface of wafer 3700.

A variety of process gases are flowed at carefully controlled rates fromreservoirs 3708 a and 3708 b, through flow channels in elastomeric block3704, and out of orifices 3706. As a result of the precisely controlledflow of process gases above wafer 3700, solid material 3710 having anextremely uniform structure is deposited.

Precise metering of reactant gas flow rates utilizing valve and/or pumpstructures of the present invention is possible for several reasons.First, gases can be flowed through valves that respond in a linearfashion to an applied actuation pressure, as is discussed above inconnection with FIGS. 21A and 21B. Alternatively or in addition tometering of gas flow using valves, the predictable behavior of pumpstructures in accordance with the present invention can be used toprecisely meter process gas flow.

In addition to the chemical vapor deposition processes described above,the present technique is also useful to control gas flow in techniquessuch as molecular beam epitaxy and reactive ion etching.

3. Micro Mirror Arrays

While the embodiments of the present invention described thus far relateto operation of a structure composed entirely of elastomeric material,the present invention is not limited to this type of structure.Specifically, it is within the scope of the present invention to combinean elastomeric structure with a conventional, silicon-basedsemiconductor structure.

For example, further contemplated uses of the present microfabricatedpumps and valves are in optical displays in which the membrane in anelastomeric structure reflects light either as a flat planar or as acurved surface depending upon whether the membrane is activated. Assuch, the membrane acts as a switchable pixel. An array of suchswitchable pixels, with appropriate control circuitry, could be employedas a digital or analog micro mirror array.

Accordingly, FIG. 38 shows an exploded view of a portion of oneembodiment of a micro mirror array in accordance with the presentinvention.

Micro mirror array 3800 includes first elastomer layer 3802 overlyingand separated from and underlying semiconductor structure 3804 by secondelastomer layer 3806. Surface 3804 a of semiconductor structure 3804bears a plurality of electrodes 3810. Electrodes 3810 are individuallyaddressable through conducting row and column lines, as would be knownto one of ordinary skill in the art.

First elastomeric layer 3802 includes a plurality of intersectingchannels 3822 underlying an electrically conducting, reflectingelastomeric membrane portion 3802 a. First elastomeric layer 3802 isaligned over second elastomeric layer 3806 and underlying semiconductordevice 3804 such that points of intersection of channels 3822 overlieelectrodes 3810.

In one embodiment of a method of fabrication in accordance with thepresent invention, first elastomeric layer 3822 may be formed byspincoating elastomeric material onto a mold featuring intersectingchannels, curing the elastomer, removing the shaped elastomer from themold, and introducing electrically conducting dopant into surface regionof the shaped elastomer. Alternatively as described in connection withFIGS. 7C-7G above, first elastomeric layer 3822 may be formed from twolayers of elastomer by inserting elastomeric material into a moldcontaining intersecting channels such that the elastomeric material isflush with the height of the channel walls, and then bonding a separatedoped elastomer layer to the existing elastomeric material to form amembrane on the top surface.

Alternatively, the first elastomeric layer 3802 may be produced fromelectrically conductive elastomer, where the electrical conductivity isdue either to doping or to the intrinsic properties of the elastomermaterial.

During operation of reflecting structure 3800, electrical signals arecommunicated along a selected row line and column line to electrode 3810a. Application of voltage to electrode 3810 a generates an attractiveforce between electrode 3810 a and overlying membrane 3802 a. Thisattractive force actuates a portion of membrane 3802 a, causing thismembrane portion to flex downward into the cavity resulting fromintersection of the channels 3822. As a result of distortion of membrane3802 a from planar to concave, light is reflected differently at thispoint in the surface of elastomer structure 3802 than from thesurrounding planar membrane surface. A pixel image is thereby created.

The appearance of this pixel image is variable, and may be controlled byaltering the magnitude of voltage applied to the electrode. A highervoltage applied to the electrode will increase the attractive force onthe membrane portion, causing further distortion in its shape. A lowervoltage applied to the electrode will decrease the attractive force onthe membrane, reducing distortion in its shape from the planar. Eitherof these changes will affect the appearance of the resulting pixelimage.

A variable micro mirror array structure as described could be used in avariety of applications, including the display of images. Anotherapplication for a variable micro mirror array structure in accordancewith an embodiment of the present invention would be as a high capacityswitch for a fiber optics communications system, with each pixel capableof affecting the reflection and transfer of a component of an incidentlight signal.

While the above embodiment describes a composite,elastomer/semiconductor structure that is utilized as a micromirrorarray, the present invention is not limited to this particularembodiment.

4. Refracting Structures

The micro-mirror array structure just described controls reflection ofincident light. However, the present invention is not limited tocontrolling reflection. Yet another embodiment of the present inventionenables the exercise of precise control over refraction of incidentlight in order to create lens and filter structures.

FIG. 39 shows one embodiment of a refractive structure in accordancewith the present invention. Refractive structure 3900 includes firstelastomeric layer 3902 and second elastomeric layer 3904 composed ofelastomeric material capable of transmitting incident light 3906.

First elastomeric layer 3902 has convex portion 3902 a which may becreated by curing elastomeric material formed over a micromachined moldhaving a concave portion. Second elastomeric layer 3904 has a flowchannel 3905 and may be created from a micromachined mold having araised line as discussed extensively above.

First elastomer layer 3902 is bonded to second elastomer layer 3904 suchthat convex portion 3902 a is positioned above flow channel 3905. Thisstructure can serve a variety of purposes.

For example, light incident to elastomeric structure 3900 would befocused into the underlying flow channel, allowing the possibleconduction of light through the flow channel. Alternatively, in oneembodiment of an elastomeric device in accordance with the presentinvention, fluorescent or phosphorescent liquid could be flowed throughthe flow channel, with the resulting light from the fluid refracted bythe curved surface to form a display.

FIG. 40 shows another embodiment of a refractive structure in accordancewith the present invention. Refractive structure 4000 is a multilayeroptical train based upon a Fresnel lens design. Specifically, refractivestructure 4000 is composed of four successive elastomer layers 4002,4004, 4006, and 4008, bonded together. The upper surfaces of each offirst, second, and third elastomer layers 4002, 4004, and 4006 bearuniform serrations 4010 regularly spaced by a distance X that is muchlarger than the wavelength of the incident light. Serrations 4010 serveto focus the incident light, and may be formed through use of amicromachined mold as described extensively above. First, second, andthird elastomer layers 4002, 4004, and 4006 function as Fresnel lensesas would be understood of one of ordinary skill in the art.

Fourth elastomeric layer 4008 bears uniform serrations 4012 having amuch smaller size than the serrations of the overlying elastomericlayers. Serrations 4012 are also spaced apart by a much smaller distanceY than the serrations of the overlying elastomeric layers, with Y on theorder of the wavelength of incident light, such that elastomeric layer4008 functions as a diffraction grating.

FIG. 41 illustrates an embodiment of a refractive structure inaccordance with the present invention which utilizes difference inmaterial refractive index to primarily accomplish diffraction.Refractive structure 4100 includes lower elastomeric portion 4102covered by upper elastomeric portion 4104. Both lower elastomericportion 4102 and upper elastomeric portion 4104 are composed of materialtransmitting incident light 4106. Lower elastomeric portion 4102includes a plurality of serpentine flow channels 4108 separated byelastomeric lands 4110. Flow channels 4108 include fluid 4112 having adifferent refractive index than the elastomeric material making up lands4110. Fluid 4112 is pumped through serpentine flow channels 4108 by theoperation of pump structure 4114 made up of parallel control channels4116 a and 4116 b overlying and separated from inlet portion 4108 a offlow channel 4108 by moveable membrane 4118.

Light 4106 incident to refractive structure 4100 encounters a series ofuniformly-spaced fluid-filled flow channels 4108 separated byelastomeric lands 4110. As a result of the differing optical propertiesof material present in these respective fluid/elastomer regions,portions of the incident light are not uniformly refracted and interactto form an interference pattern. A stack of refractive structures of themanner just described can accomplish even more complex and specializedrefraction of incident light.

The refractive elastomeric structures just described can fulfill avariety of purposes. For example, the elastomeric structure could act asa filter or optical switch to block selected wavelengths of incidentlight. Moreover, the refractive properties of the structure could bereadily adjusted depending upon the needs of a particular application.

For example, the composition (and hence refractive index) of fluidflowed through the flow channels could be changed to affect diffraction.Alternatively, or in conjunction with changing the identity of the fluidflowed, the distance separating adjacent flow channels can be preciselycontrolled during fabrication of the structure in order to generate anoptical interference pattern having the desired characteristics.

5. Normally-Closed Valve Structure

FIGS. 7B and 7H above depict a valve structure in which the elastomericmembrane is moveable from a first relaxed position to a second actuatedposition in which the flow channel is blocked. However, the presentinvention is not limited to this particular valve configuration.

FIGS. 42A-42J show a variety of views of a normally-closed valvestructure in which the elastomeric membrane is moveable from a firstrelaxed position blocking a flow channel, to a second actuated positionin which the flow channel is open, utilizing a negative controlpressure.

FIG. 42A shows a plan view, and FIG. 42B shows a cross sectional viewalong line 42B-42B′, of normally-closed valve 4200 in an unactuatedstate. Flow channel 4202 and control channel 4204 are formed inelastomeric block 4206 overlying substrate 4205. Flow channel 4202includes a first portion 4202 a and a second portion 4202 b separated byseparating portion 4208. Control channel 4204 overlies separatingportion 4208. As shown in FIG. 42B, in its relaxed, unactuated position,separating portion 4008 remains positioned between flow channel portions4202 a and 4202 b, interrupting flow channel 4202.

FIG. 42C shows a cross-sectional view of valve 4200 wherein separatingportion 4208 is in an actuated position. When the pressure withincontrol channel 4204 is reduced to below the pressure in the flowchannel (for example by vacuum pump), separating portion 4208experiences an actuating force drawing it into control channel 4204. Asa result of this actuation force membrane 4208 projects into controlchannel 4204, thereby removing the obstacle to a flow of materialthrough flow channel 4202 and creating a passageway 4203. Upon elevationof pressure within control channel 4204, separating portion 4208 willassume its natural position, relaxing back into and obstructing flowchannel 4202.

The behavior of the membrane in response to an actuation force may bechanged by varying the width of the overlying control channel.Accordingly, FIGS. 42D-42H show plan and cross-sectional views of analternative embodiment of a normally-closed valve 4201 in which controlchannel 4207 is substantially wider than separating portion 4208. Asshown in cross-sectional views FIG. 42E-F along line 42E-42E′ of FIG.42D, because a larger area of elastomeric material is required to bemoved during actuation, the actuation force necessary to be applied isreduced.

FIGS. 42G and H show a cross-sectional views along line 40G-40G′ of FIG.40D. In comparison with the unactuated valve configuration shown in FIG.42G, FIG. 42H shows that reduced pressure within wider control channel4207 may under certain circumstances have the unwanted effect of pullingunderlying elastomer 4206 away from substrate 4205, thereby creatingundesirable void 4212.

Accordingly, FIG. 42I shows a plan view, and 42J a cross-sectional viewalong line 42J-42J′ of FIG. 42I, of valve structure 4220 which avoidsthis problem by featuring control line 4204 with a minimum width exceptin segment 4204 a overlapping separating portion 4208. As shown in FIG.42J, even under actuated conditions the narrower cross-section ofcontrol channel 4204 reduces the attractive force on the underlyingelastomer material 4206, thereby preventing this elastomer material frombeing drawn away from substrate 4205 and creating an undesirable void.

While a normally-closed valve structure actuated in response to pressureis shown in FIGS. 42A-42J, a normally-closed valve in accordance withthe present invention is not limited to this configuration. For example,the separating portion obstructing the flow channel could alternativelybe manipulated by electric or magnetic fields, as described extensivelyabove.

6. Separation of Materials

In a further application of the present invention, an elastomericstructure can be utilized to perform separation of materials. FIG. 43shows one embodiment of such a device.

Separation device 4300 features an elastomeric block 4301 includingfluid reservoir 4302 in communication with flow channel 4304. Fluid ispumped from fluid reservoir 4306 through flow channel 4308 byperistaltic pump structure 4310 formed by control channels 4312overlying flow channel 4304, as has been previously described at length.Alternatively, where a peristaltic pump structure in accordance with thepresent invention is unable to provide sufficient back pressure, fluidfrom a reservoir positioned outside the elastomeric structure may bepumped into the elastomeric device utilizing an external pump.

Flow channel 4304 leads to separation column 4314 in the form of achannel packed with separation matrix 4316 behind porous frit 4318. Asis well known in the art of chromatography, the composition of theseparation matrix 4316 depends upon the nature of the materials to beseparated and the particular chromatography technique employed. Theelastomeric separation structure is suitable for use with a variety ofchromatographic techniques, including but not limited to gel exclusion,gel permeation, ion exchange, reverse phase, hydrophobic interaction,affinity chromatography, fast protein liquid chromatography (FPLC) andall formats of high pressure liquid chromatography (HPLC). The highpressures utilized for HPLC may require the use of urethane,dicyclopentadiene or other elastomer combinations.

Samples are introduced into the flow of fluid into separation column4314 utilizing load channel 4319. Load channel 4319 receives fluidpumped from sample reservoir 4320 through pump 4321. Upon opening ofvalve 4322 and operation of pump 4321, sample is flowed from loadchannel 4319 into flow channel 4304. The sample is then flowed throughseparation column 4314 by the action of pump structure 4312. As a resultof differential mobility of the various sample components in separationmatrix 4316, these sample components become separated and are elutedfrom column 4314 at different times.

Upon elution from separation column 4314, the various sample componentspass into detection region 4324. As is well known in the art ofchromatography, the identity of materials eluted into detection region4324 can be determined utilizing a variety of techniques, including butnot limited to fluorescence, UV/visible/IR spectroscopy, radioactivelabeling, amperometric detection, mass spectroscopy, and nuclearmagnetic resonance (NMR).

A separation device in accordance with the present invention offers theadvantage of extremely small size, such that only small volumes of fluidand sample are consumed during the separation. In addition, the deviceoffers the advantage of increased sensitivity. In conventionalseparation devices, the size of the sample loop will prolong theinjection of the sample onto the column, causing width of the elutedpeaks to potentially overlap with one another. The extremely small sizeand capacity of the load channel in general prevents this peak diffusionbehavior from becoming a problem.

The separation structure shown in FIG. 43 represents only one embodimentof such a device, and other structures are contemplated by the presentinvention. For example, while the separation device of FIG. 43 featuresa flow channel, load loop, and separation column oriented in a singleplane, this is not required by the present invention. One or more of thefluid reservoir, the sample reservoir, the flow channel, the load loop,and the separation column could be oriented perpendicular to one anotherand/or to the plane of the elastomeric material utilizing via structureswhose formation is described at length above in connection with FIG.35A-D.

7. Cell Pen/Cell Cage/Cell Grinder

In yet a further application of the present invention, an elastomericstructure can be utilized to manipulate organisms or other biologicalmaterial. FIGS. 44A-44D show plan views of one embodiment of a cell penstructure in accordance with the present invention.

Cell pen array 4400 features an array of orthogonally-oriented flowchannels 4402, with an enlarged “pen” structure 4404 at the intersectionof alternating flow channels. Valve 4406 is positioned at the entranceand exit of each pen structure 4404. Peristaltic pump structures 4408are positioned on each horizontal flow channel and on the vertical flowchannels lacking a cell pen structure.

Cell pen array 4400 of FIG. 44A has been loaded with cells A-H that havebeen previously sorted, perhaps by a sorting structure as describedabove in conjunction with FIG. 36. FIGS. 44B-44C show the accessing andremoval of individually stored cell C by 1) opening valves 4406 oneither side of adjacent pens 4404 a and 4404 b, 2) pumping horizontalflow channel 4402 a to displace cells C and G, and then 3) pumpingvertical flow channel 4402 b to remove cell C. FIG. 44D shows thatsecond cell G is moved back into its prior position in cell pen array4400 by reversing the direction of liquid flow through horizontal flowchannel 4402 a.

The cell pen array 4404 described above is capable of storing materialswithin a selected, addressable position for ready access. However,living organisms such as cells may require a continuous intake of foodsand expulsion of wastes in order to remain viable. Accordingly, FIGS.45A and 45B show plan and cross-sectional views (along line 45B-45B′)respectively, of one embodiment of a cell cage structure in accordancewith the present invention.

Cell cage 4500 is formed as an enlarged portion 4500 a of a flow channel4501 in an elastomeric block 4503 in contact with substrate 4505. Cellcage 4500 is similar to an individual cell pen as described above inFIGS. 44A-44D, except that ends 4500 b and 4500 c of cell cage 4500 donot completely enclose interior region 4500 a. Rather, ends 4500 a and4500 b of cage 4500 are formed by a plurality of retractable pillars4502. Pillars 4502 may be part of a membrane structure of anormally-closed valve structure as described extensively above inconnection with FIGS. 42A-42J.

Specifically, control channel 4504 overlies pillars 4502. When thepressure in control channel 4504 is reduced, elastomeric pillars 4502are drawn upward into control channel 4504, thereby opening end 4500 bof cell cage 4500 and permitting a cell to enter. Upon elevation ofpressure in control channel 4504, pillars 4502 relax downward againstsubstrate 4505 and prevent a cell from exiting cage 4500.

Elastomeric pillars 4502 are of a sufficient size and number to preventmovement of a cell out of cage 4500, but also include gaps 4508 whichallow the flow of nutrients into cage interior 4500 a in order tosustain cell(s) stored therein. Pillars 4502 on opposite end 4500 c aresimilarly configured beneath second control channel 4506 to permitopening of the cage and removal of the cell as desired.

Under certain circumstances, it may be desirable to grind/disrupt cellsor other biological materials in order to access component pieces.

Accordingly, FIGS. 46A and 46B show plan and cross sectional views(along line 46B-46B′) respectively, of one embodiment of cell grinderstructure 4600 in accordance with the present invention. Cell grinder4600 includes a system of interdigitated posts 4602 within flow channel4604 which close together upon actuation of integral membrane 4606 byoverlying control channel 4608. By closing together, posts 4602 crushmaterial present between them.

Posts 4602 may be spaced at intervals appropriate to disrupt entities(cells) of a given size. For disruption of cellular material, spacing ofposts 4602 at an interval of about 2 μm is appropriate. In alternativeembodiments of a cell grinding structure in accordance with the presentinvention, posts 4602 may be located entirely on the above-lyingmembrane, or entirely on the floor of the control channel.

8. Pressure Oscillator

In yet a further application of the present invention, an elastomericstructure can be utilized to create a pressure oscillator structureanalogous to oscillator circuits frequently employed in the field ofelectronics. FIG. 47 shows a plan view of one embodiment of such apressure oscillator structure.

Pressure oscillator 4700 comprises an elastomeric block 4702 featuringflow channel 4704 formed therein. Flow channel 4704 includes an initialportion 4704 a proximate to pressure source 4706, and a serpentineportion 4704 b distal from pressure source 4706. Initial portion 4704 ais in contact with via 4708 in fluid communication with control channel4710 formed in elastomeric block 4702 above the level of flow channel4704. At a location more distal from pressure source 4706 than via 4708,control channel 4710 overlies and is separated from flow channel 4704 byan elastomeric membrane, thereby forming valve 4712 as previouslydescribed.

Pressure oscillator structure 4700 operates as follows. Initially,pressure source 4706 provides pressure along flow channel 4704 andcontrol channel 4710 through via 4708. Because of the serpentine shapeof flow channel 4704 b, pressure is lower in region 4704 b as comparedwith flow channel 4710. At valve 4712, the pressure difference betweenserpentine flow channel portion 4704 b and overlying control channel4710 eventually causes the membrane of valve 4712 to project downwardinto serpentine flow channel portion 4704 b, closing valve 4712. Owingto the continued operation of pressure source 4706 however, pressurebegins to build up in serpentine flow channel portion 4704 b behindclosed valve 4712. Eventually the pressure equalizes between controlchannel 4710 and serpentine flow channel portion 4704 b, and valve 4712opens.

Given the continuous operation of the pressure source, theabove-described build up and release of pressure will continueindefinitely, resulting in a regular oscillation of pressure. Such apressure oscillation device may perform any number of possiblefunctions, including but not limited to timing.

9. Side-Actuated Valve

While the above description has focused upon microfabricated elastomericvalve structures in which a control channel is positioned above andseparated by an intervening elastomeric membrane from an underlying flowchannel, the present invention is not limited to this configuration.FIGS. 48A and 48B show plan views of one embodiment of a side-actuatedvalve structure in accordance with one embodiment of the presentinvention.

FIG. 48A shows side-actuated valve structure 4800 in an unactuatedposition.

Flow channel 4802 is formed in elastomeric layer 4804. Control channel4806 abutting flow channel 4802 is also formed in elastomeric layer4804. Control channel 4806 is separated from flow channel 4802 byelastomeric membrane portion 4808. A second elastomeric layer (notshown) is bonded over bottom elastomeric layer 4804 to enclose flowchannel 4802 and control channel 4806.

FIG. 48B shows side-actuated valve structure 4800 in an actuatedposition. In response to a build up of pressure within control channel4806, membrane 4808 deforms into flow channel 4802, blocking flowchannel 4802. Upon release of pressure within control channel 4806,membrane 4808 would relax back into control channel 4806 and open flowchannel 4802.

While a side-actuated valve structure actuated in response to pressureis shown in FIGS. 48A and 48B, a side-actuated valve in accordance withthe present invention is not limited to this configuration. For example,the elastomeric membrane portion located between the abutting flow andcontrol channels could alternatively be manipulated by electric ormagnetic fields, as described extensively above.

10. Additional Applications

The following represent further aspects of the present invention:present valves and pumps can be used for drug delivery (for example, inan implantable drug delivery device); and for sampling of biologicalfluids (for example, by storing samples sequentially in a column withplugs of spacer fluid therebetween, wherein the samples can be shuntedinto different storage reservoirs, or passed directly to appropriatesensor(s). Such a fluid sampling device could also be implanted in thepatient's body.

The present systems can also be used for devices which relieveover-pressure in vivo using a micro-valve or pump. For example, animplantable bio-compatible micro-valve can be used to relieveover-pressures in the eye which result from glaucoma. Other contemplateduses of the present switchable micro-valves include implantation in thespermatic duct or fallopian tube allowing reversible long-term orshort-term birth control without the use of drugs.

Further uses of the present invention include DNA sequencing whereby theDNA to be sequenced is provided with a polymerase and a primer, and isthen exposed to one type of DNA base (A, C, T, or G) at a time in orderto rapidly assay for base incorporation. In such a system, the basesmust be flowed into the system and excess bases washed away rapidly.Pressure driven flow, gated by elastomeric micro-valves in accordancewith the present invention would be ideally suited to allow for suchrapid flow and washing of reagents.

Other contemplated uses of the present micro-valve and micro-pumpsystems include uses with DNA chips. For example, a sample can be flowedinto a looped channel and pumped around the loop with a peristalticaction such that the sample can make many passes over the probes of theDNA array. Such a device would give the sample that would normally bewasted sitting over the non-complimentary probes the chance to bind to acomplimentary probe instead. An advantage of such a looped-flow systemis that it would reduce the necessary sample volume, and therebyincrease assay sensitivity.

Further applications exist in high throughput screening in whichapplications could benefit by the dispensing of small volumes of liquid,or by bead-based assays wherein ultrasensitive detection wouldsubstantially improve assay sensitivity.

Another contemplated application is the deposition of array of variouschemicals, especially oligonucleotides, which may or may not have beenchemically fabricated in a previous action of the device beforedeposition in a pattern or array on a substrate via contact printingthrough fluid channel outlets in the elastomeric device in closeproximity to a desired substrate, or by a process analogous to ink jetprinting.

The present microfabricated elastomeric valves and pumps could also beused to construct systems for reagent dispensing, mixing and reactionfor synthesis of oligonucleotides, peptides or other biopolymers.

Further applications for the present invention include ink jet printerheads, in which small apertures are used to generate a pressure pulsesufficient to expel a droplet. An appropriately actuated micro-valve inaccordance with the present invention can create such a pressure pulse.The present micro-valves and pumps can also be used to digitallydispense ink or pigment, in amounts not necessarily as small as singledroplets. The droplet would be brought into contact with the mediumbeing printed on rather than be required to be fired through the air.

Yet other uses of the present systems are in fluidic logic circuitswhich offer the advantages of being useable in radiation resistantapplications. A further advantage of such fluidic logic circuits isthat, being non-electronic, such fluidic logic circuitry may not beprobed by electro magnetic sensors, thus offering a security benefit.

Yet further uses of the present invention would take advantage of theready removal and reattachment of the structure from an underlyingsubstrate such as glass, utilizing a glass substrate patterned with abinding or other material. This allows separate construction of apatterned substrate and elastomer structure. For instance, a glasssubstrate could be patterned with a DNA microarray, and an elastomervalve and pump structure sealed over the array in a subsequent step.

11. Additional Aspects of the Invention

The following represent further aspects of the present invention: theuse of a deflectable membrane to control flow of a fluid in amicrofabricated channel of an elastomeric structure; the use ofelastomeric layers to make a microfabricated elastomeric devicecontaining a microfabricated movable portion; and the use of anelastomeric material to make a microfabricated valve or pump.

A microfabricated elastomeric structure in accordance with oneembodiment of the present invention comprises an elastomeric blockformed with microfabricated recesses therein, a portion of theelastomeric block deflectable when the portion is actuated. The recessescomprise a first microfabricated channel and a first microfabricatedrecess, and the portion comprises an elastomeric membrane deflectableinto the first microfabricated channel when the membrane is actuated.The recesses have a width in the range of 10 μm to 200 μm and theportion has a thickness of between about 2 μm and 50 μm. Themicrofabricated elastomeric structure may be actuated at a speed of 100Hz or greater and contains substantially no dead volume when the portionis actuated.

A method of actuating an elastomeric structure comprises providing anelastomeric block formed with first and second microfabricated recessestherein, the first and second microfabricated recesses separated by amembrane portion of the elastomeric block deflectable into one of thefirst and second recesses in response to an actuation force, andapplying an actuation force to the membrane portion such that themembrane portion is deflected into one of the first and the secondrecesses.

A method of microfabricating an elastomeric structure in accordance withone embodiment of the present invention comprises forming a firstelastomeric layer on a substrate, curing the first elastomeric layer,and patterning a first sacrificial layer over the first elastomericlayer. A second elastomeric layer is formed over the first elastomericlayer, thereby encapsulating the first patterned sacrificial layerbetween the first and second elastomeric layers, the second elastomericlayer is cured, and the first patterned sacrificial layer is removedselective to the first elastomeric layer and the second elastomericlayer, thereby forming at least one first recess between the first andsecond layers of elastomer.

An alternative embodiment of a method of fabricating further comprisespatterning a second sacrificial layer over the substrate prior toforming the first elastomeric layer, such that the second patternedsacrificial layer is removed during removal of the first patternedsacrificial layer to form at least one recess along a bottom of thefirst elastomeric layer.

A microfabricated elastomeric structure in accordance with oneembodiment of the present invention comprises an elastomeric block, afirst channel and a second channel separated by a separating portion ofthe elastomeric structure, and a microfabricated recess in theelastomeric block adjacent to the separating portion such that theseparating portion may be actuated to deflect into the microfabricatedrecess. 66. Deflection of the separating portion opens a passagewaybetween the first and second channels.

A method of controlling fluid or gas flow through an elastomericstructure comprises providing an elastomeric block, the elastomericblock having first, second, and third microfabricated recesses, and theelastomeric block having a first microfabricated channel passingtherethrough, the first, second and third microfabricated recessesseparated from the first channel by respective first, second and thirdmembranes deflectable into the first channel, and deflecting the first,second and third membranes into the first channel in a repeatingsequence to peristaltically pump a flow of fluid through the firstchannel.

A method of microfabricating an elastomeric structure comprisesmicrofabricating a first elastomeric layer, microfabricating a secondelastomeric layer; positioning the second elastomeric layer on top ofthe first elastomeric layer; and bonding a bottom surface of the secondelastomeric layer onto a top surface of the first elastomeric layer.

12. Alternative Device Fabrication Methods

FIGS. 1-7A and 8-18 above show embodiments of multilayer softlithography and encapsulation methods, respectively, for fabricatingelastomer structures in accordance with the present invention. However,these fabrication methods are merely exemplary, and variations on thesetechniques may be employed to create elastomer structures.

For example, FIGS. 3 and 4 show fabrication of an elastomer structureutilizing multilayer soft lithography techniques, wherein therecess-bearing face of the upper elastomer layer is placed on top of thenon-recess-bearing face of the lower elastomer layer. However, thepresent invention is not limited to this configuration.

FIG. 50 shows elastomeric structure 5410 featuring an alternativeorientation of elastomeric layers, wherein recess-bearing faces 5400 and5402 of first and second elastomer layers 5404 and 5406 are respectivelyplaced into contact to create a larger-sized channel 5408.Alternatively, FIG. 51 shows an orientation of elastomeric layerswherein non-recess bearing faces 5500 and 5502 are placed into contact,such that recesses are present on opposite sides of the structure. Suchan elastomer structure 5512 could be sandwiched between substrates 5508and 5510 to produce channels 5504 and 5506 that cross-over each other.

FIGS. 52A-52D show various views of steps for constructing a fluidchannel bridging structure utilizing encapsulation methods.Specifically, FIG. 52A shows a cross-sectional view of the formation oflower elastomeric portion 5600 of bridging structure 5602. FIG. 52Bshows a cross-sectional view of the formation of upper elastomericportion 5604 of the bridging structure. FIG. 52C shows assembly of upperand lower elastomeric portions 5604 and 5600 to form bridging structure5602, wherein fluid flowing in channel 5606 bridges over cross-channel5608 through vias 5610 and 5612 and bridge portion 5614 as shown by thearrows. FIG. 52D shows a plan view of bridging structure 5602, withbridge portion 5614 of the upper elastomeric is shown in solid, and thefeatures in the underlying lower elastomer layer are shown in outline.This structure requires the formation of vias in the lower layer, butallows multiple liquid streams to flow in a single layer and cross overone another without mixing.

13. Composite Structures

As discussed above in conjunction with the micromirror array structureof FIG. 38, the fabricated elastomeric structures of the presentinvention may be combined with non-elastomeric materials to createcomposite structures. Fabrication of such composite structures is nowdiscussed in further detail.

FIG. 53 shows a cross-sectional view of one embodiment of a compositestructure in accordance with the present invention. FIG. 53 showscomposite valve structure 5700 including first, thin elastomer layer5702 overlying semiconductor-type substrate 5704 having channel 5706formed therein. Second, thicker elastomer layer 5708 overlies firstelastomer layer 5702. Actuation of first elastomer layer 5702 to driveit into channel 5706, will cause composite structure 5700 to operate asa valve.

FIG. 54 shows a cross-sectional view of a variation on this theme,wherein thin elastomer layer 5802 is sandwiched between two hard,semiconductor substrates 5804 and 5806, with lower substrate 5804featuring channel 5808. Again, actuation of thin elastomer layer 5802 todrive it into channel 5808 will cause composite structure 5810 tooperate as a valve.

The structures shown in FIG. 53 or 54 may be fabricated utilizing eitherthe multilayer soft lithography or encapsulation techniques describedabove. In the multilayer soft lithography method, the elastomer layer(s)would be formed and then placed over the semiconductor substrate bearingthe channel. In the encapsulation method, the channel would be firstformed in the semiconductor substrate, and then the channel would befilled with a sacrificial material such as photoresist. The elastomerwould then be formed in place over the substrate, with removal of thesacrificial material producing the channel overlaid by the elastomermembrane. As is discussed in detail below in connection with bonding ofelastomer to other types of materials, the encapsulation approach mayresult in a stronger seal between the elastomer membrane component andthe underlying nonelastomer substrate component.

As shown in FIGS. 53 and 54, a composite structure in accordance withembodiments of the present invention may include a hard substrate thatbears a passive feature such as a channels. However, the presentinvention is not limited to this approach, and the underlying hardsubstrate may bear active features that interact with an elastomercomponent bearing a recess. This is shown in FIG. 55, wherein compositestructure 5900 includes elastomer component 5902 containing recess 5904having walls 5906 and ceiling 5908. Ceiling 5908 forms flexible membraneportion 5909. Elastomer component 5902 is sealed against substantiallyplanar nonelastomeric component 5910 that includes active device 5912.Active device 5912 may interact with material present in recess 5904and/or flexible membrane portion 5909.

In the micromirror array embodiment previously described in conjunctionwith FIG. 38, the underlying substrate contained active structures inthe form of an electrode array. However, many other types of activestructures may be present in the nonelastomer substrate. Activestructures that could be present in an underlying hard substrateinclude, but are not limited to, resistors, capacitors, photodiodes,transistors, chemical field effect transistors (chem FET's),amperometric/coulometric electrochemical sensors, fiber optics, fiberoptic interconnects, light emitting diodes, laser diodes, verticalcavity surface emitting lasers (VCSEL's), micromirrors, accelerometers,pressure sensors, flow sensors, CMOS imaging arrays, CCD cameras,electronic logic, microprocessors, thermistors, Peltier coolers,waveguides, resistive heaters, chemical sensors, strain gauges,inductors, actuators (including electrostatic, magnetic,electromagnetic, bimetallic, piezoelectric, shape-memory-alloy based,and others), coils, magnets, electromagnets, magnetic sensors (such asthose used in hard drives, superconducting quantum interference devices(SQUIDS) and other types), radio frequency sources and receivers,microwave frequency sources and receivers, sources and receivers forother regions of the electromagnetic spectrum, radioactive particlecounters, and electrometers.

As is well known in the art, a vast variety of technologies can beutilized to fabricate active features in semiconductor and other typesof hard substrates, including but not limited printed circuit board(PCB) technology, CMOS, surface micromachining, bulk micromachining,printable polymer electronics, and TFT and otheramorphous/polycrystalline techniques as are employed to fabricate laptopand flat screen displays.

A composite structure in accordance with one embodiment of the presentinvention comprises a nonelastomer substrate having a surface bearing afirst recess, a flexible elastomer membrane overlying the non-elastomersubstrate, the membrane able to be actuated into the first recess, and alayer overlying the flexible elastomer membrane.

An embodiment of a method of forming a composite structure comprisesforming a recess in a first nonelastomer substrate, filling the recesswith a sacrificial material, forming a thin coat of elastomer materialover the nonelastomer substrate and the filled recess, curing theelastomer to form a thin membrane, and removing the sacrifical material.

A composite structure in accordance with an alternative embodiment ofthe present invention comprises an elastomer component defining a recesshaving walls and a ceiling, the ceiling of the recess forming a flexiblemembrane portion, and a substantially planar nonelastomer componentsealed against the elastomer component, the nonelastomer componentincluding an active device interacting with at least one of the membraneportion and a material present in the recess.

A method of fabricating a composite structure comprising forming arecess in an elastomer component, the recess including walls and aceiling, the ceiling forming a flexible membrane portion, positioning amaterial within the recess, forming a substantially planar nonelastomercomponent including an active device, and sealing the elastomercomponent against the nonelastomer component, such that the activedevice may interact with at least one of the membrane portion and thematerial.

A variety of approaches can be employed to seal the elastomericstructure against the nonelastomeric substrate, ranging from thecreation of a Van der Waals bond between the elastomeric andnonelastomeric components, to creation of covalent or ionic bondsbetween the elastomeric and nonelastomeric components of the compositestructure. Example approaches to sealing the components together arediscussed below, approximately in order of increasing strength.

A first approach is to rely upon the simple hermetic seal resulting fromVan der Waals bonds formed when a substantially planar elastomer layeris placed into contact with a substantially planar layer of a harder,non-elastomer material. In one embodiment, bonding of RTV elastomer to aglass substrate created a composite structure capable of withstanding upto about 3-4 psi of pressure. This may be sufficient for many potentialapplications.

A second approach is to utilize a liquid layer to assist in bonding. Oneexample of this involves bonding elastomer to a hard glass substrate,wherein a weakly acidic solution (5 μl HCl in H₂0, pH 2) was applied toa glass substrate. The elastomer component was then placed into contactwith the glass substrate, and the composite structure baked at 37° C. toremove the water. This resulted in a bond between elastomer andnon-elastomer able to withstand a pressure of about 20 psi. In thiscase, the acid may neutralize silanol groups present on the glasssurface, permitting the elastomer and non-elastomer to enter into goodVan der Waals contact with each other.

A third approach is to create a covalent chemical bond between theelastomer component and functional groups introduced onto the surface ofa nonelastomer component. Examples of derivitization of a nonelastomersubstrate surface to produce such functional groups include exposing aglass substrate to agents such as vinyl silane or aminopropyltriethoxysilane (APTES), which may be useful to allow bonding of the glass tosilicone elastomer and polyurethane elastomer materials, respectively.

A fourth approach is to create a covalent chemical bond between theelastomer component and a functional group native to the surface of thenonelastomer component. For example, RTV elastomer can be created withan excess of vinyl groups on its surface. These vinyl groups can becaused to react with corresponding functional groups present on theexterior of a hard substrate material, for example the Si—H bondsprevalent on the surface of a single crystal silicon substrate afterremoval of native oxide by etching. In this example, the strength of thebond created between the elastomer component and the nonelastomercomponent has been observed to exceed the materials strength of theelastomer components.

While the present invention has been described herein with reference toparticular embodiments thereof, a latitude of modification, variouschanges and substitutions are intended in the foregoing disclosure, andit will be appreciated that in some instances some features of theinvention will be employed without a corresponding use of other featureswithout departing from the scope of the invention as set forth.Therefore, many modifications may be made to adapt a particularsituation or material to the teachings of the invention withoutdeparting from the essential scope and spirit of the present invention.It is intended that the invention not be limited to the particularembodiment disclosed as the best mode contemplated for carrying out thisinvention, but that the invention will include all embodiments andequivalents falling within the scope of the claims.

The following pending patent applications contain subject matter relatedto the instant application and are hereby incorporated by reference:U.S. nonprovisional patent application Ser. No. ______, entitled“Apparatus and Methods for Conducting Cell Assays and High ThroughputScreening” (Atty. docket No. 009195-000200US), filed Nov. 16, 2000; U.S.nonprovisional patent application Ser. No. ______, entitled “IntegratedActive Flux Microfluidic Devices and Methods” (Atty docket no.0G638-US1), filed Nov. 16, 2000; and U.S. nonprovisional patentapplication Ser. No. ______, entitled “Fabrication of Micro and NanoScale Devices With Soft Materials” (Atty. docket No. OH944), filed Nov.16, 2000.

“Monolithic Microfabricated Valves and Pumps by Multilayer SoftLithography,” Unger et al., Science, Vol. 288, pp. 113-116 (Apr. 7,2000), which is herein incorporated by reference in its entirety for allpurposes.

1-17. (canceled)
 18. A microfabricated elastomeric device comprising: aplanar substrate; a first elastomeric layer having a top surface and abottom surface, the bottom surface comprising a first recess, whereinthe bottom surface of the first elastomeric layer is bonded to theplanar substrate such that the first recess forms a flow channel; and asecond elastomeric layer having a top surface and a bottom surface, thebottom surface comprising a second recess, wherein the bottom surface ofthe second elastomeric layer is bonded to the top surface of the firstelastomeric layer such that the second recess forms a flow channel,wherein the second flow channel extends across and above a first flowchannel, and wherein a portion of the first elastomeric layer is adeflectable portion and is deflectable into the first flow channel uponpressurization of the second flow channel to form a planar sealingsurface between the planar substrate and the deflectable portion of thefirst elastomeric layer, and wherein the deflectable portion ischaracterized by a dead volume.
 19. The microfabricated elastomericdevice of claim 18, wherein the first and second flow channels both passthrough the elastomeric structure.
 20. The microfabricated elastomericdevice of claim 18, wherein the second flow channel passes through theelastomeric structure and the first flow channel passes along a surfaceof the elastomeric structure.
 21. The microfabricated elastomeric deviceof claim 18 wherein the first and second flow channels have a depthbetween 0.2 and 250 microns.
 22. The microfabricated elastomeric deviceof claim 18, wherein the deflectable portion is made from the samematerial as the first elastomeric layer.
 23. The microfabricatedelastomeric device of claim 18, wherein the first elastomeric layer andthe second elastomeric layer are both made form an elastomeric materialhaving a Young's modulus of between about 20 Pa-1 GPa.
 24. Themicrofabricated elastomeric device of claim 18 wherein the dead volumeis 100 aL to 100 nL.
 25. The microfabricated elastomeric device of claim18 wherein the dead volume is 1 fL to 10 nL.
 26. The microfabricatedelastomeric device of claim 18 wherein the dead volume is 100 fL to 1nL.
 27. The microfabricated elastomeric device of claim 18 wherein thedead volume is 1 pL to 100 pL.
 28. The microfabricated elastomericdevice of claim 18 wherein the dead volume is 1 aL to 1 μL.
 29. Amicrofabricated elastomeric device comprising: a planar substrate, afirst elastomeric layer having a top surface and a bottom surface, thebottom surface comprising a first recess, wherein the bottom surface ofthe first elastomeric layer is bonded to the planar substrate such thatthe first recess forms a flow channel; a second elastomeric layer havinga top surface and a bottom surface, the bottom surface comprising asecond recess, wherein the bottom surface of the second elastomericlayer is bonded to the top surface of the first elastomeric layer suchthat the second recess forms a flow channel, and wherein the second flowchannel extends across and above a first flow channel; and anelastomeric membrane bonded between the first and the second elastomericlayers, separating the first flow channel and the second flow channel,wherein the membrane is deflectable into the first flow channel uponpressurization of the second flow channel to form a planar sealingsurface between the planar substrate and the deflectable portion of thefirst elastomeric layer, and wherein the membrane is characterized by adead volume.
 30. The microfabricated elastomeric device of claim 29,wherein the first flow channel experiences a reduced pressure whichcauses the membrane to deflect into the first flow channel.
 31. Themicrofabricated elastomeric device of claim 29, wherein the first andsecond elastomeric layers comprise a material selected from the groupconsisting of: polyisoprene, polybutadiene, polychloroprene,polyisobutylene, poly(styrene-butadiene-styrene), the polyurethanes, andsilicones.
 32. The microfabricated elastomeric device of claim 29,wherein the first and second elastomeric layers comprise a materialselected from the group consisting of:poly(bis(fluoroalkoxy)phosphazene), poly(carborane-siloxanes),poly(1-butene), poly(chlorotrifluoroethylene-vinyldene fluoride)copolymers, poly(ethyl vinyl ether), poly(vinylidene fluoride),poly(vinylidene fluoride-hexafluoropropylene) copolymer.
 33. Themicrofabricated elastomeric device of claim 29, wherein the first andsecond elastomeric layers comprise a material selected from the groupconsisting of: elastomer compositions of polyvinylchloride, polysulfone,polycarbonate, polymethylmethacrylate, or polytertrafluoroethylene. 34.The microfabricated elastomeric device of claim 29, wherein the membraneis composed of an elastomeric material different than that forming thefirst or the second elastomeric layers.
 35. The microfabricatedelastomeric device of claim 29, wherein the membrane is composed of anelastomeric material the same as that forming the first or the secondelastomeric layers.
 36. The microfabricated elastomeric device of claim29 wherein the dead volume is 100 aL to 100 nL.
 37. The microfabricatedelastomeric device of claim 29 wherein the dead volume is 1 fL to 10 nL.38. The microfabricated elastomeric device of claim 29 wherein the deadvolume is 100 fL to 1 nL.
 39. The microfabricated elastomeric device ofclaim 29 wherein the dead volume is 1 pL to 100 pL.
 40. Themicrofabricated elastomeric device of claim 29 wherein the dead volumeis 1 aL to 1 μL.
 41. The microfabricated elastomeric device of claim 29wherein the first and second flow channels have a depth between 0.2 and250 microns.
 42. A microfabricated elastomeric device comprising: aplanar substrate; a first elastomeric layer having a top surface and abottom surface, the bottom surface comprising a first recess, whereinthe bottom surface of the first elastomeric layer is bonded to theplanar substrate such that the first recess forms a first flow channelwith a top and a bottom wherein the top of the first flow channel has acurved surface; and a second elastomeric layer having a top surface anda bottom surface, the bottom surface comprising a second recess, whereinthe bottom surface of the second elastomeric layer is bonded to the topsurface of the first elastomeric layer such that the second recess formsa second flow channel, wherein the second flow channel extends acrossand above a first flow channel, and wherein a portion of the firstelastomeric layer is a deflectable portion and is deflectable into thefirst flow channel upon pressurization of the second flow channel toform a planar sealing surface between the planar substrate and thedeflectable portion of the first elastomeric layer, and wherein thedeflectable portion is characterized by a dead volume.
 43. Themicrofabricated elastomeric device of claim 42 wherein the dead volumeis 100 aL to 100 nL.
 44. The microfabricated elastomeric device of claim42 wherein the dead volume is 1 fL to 10 nL.
 45. The microfabricatedelastomeric device of claim 42 wherein the dead volume is 100 fL to 1nL.
 46. The microfabricated elastomeric device of claim 42 wherein thedead volume is 1 pL to 100 pL.
 47. The microfabricated elastomericdevice of claim 42 wherein the dead volume is 1 aL to 1 μL.
 48. Themicrofabricated elastomeric device of claim 42 wherein the first andsecond flow channels have a depth between 0.2 and 250 microns.